CN122580098A

CN122580098A - Modulators of BCL6 for use in methods of treating cancer or autoimmune diseases

Info

Publication number: CN122580098A
Application number: CN202480071745.9A
Authority: CN
Inventors: D·S·莫滕森; D·黄; M·D·亚历山大; B·W·怀特菲尔德; H·P·舒纳特纳; D·S·多德; G·米西奥; N·霍尔伯格-道格拉斯; J·罗德斯; J·格里芬
Original assignee: Bristol Myers Squibb Co
Current assignee: Bristol Myers Squibb Co
Priority date: 2023-09-14
Filing date: 2024-09-13
Publication date: 2026-08-14
Also published as: WO2025059429A1; EP4777110A1; KR20260093302A

Abstract

Provided herein are compounds and compositions thereof for modulating BCL 6. In some embodiments, the compounds and compositions are provided for use in the treatment of cancer or autoimmune diseases. In some embodiments, the compound has formula (IA):

Description

Modulators of BCL6 for use in methods of treating cancer or autoimmune diseases

Cross Reference to Related Applications

The present application claims priority from U.S. provisional application No. 63/538,402 filed on 9/14 of 2023, which is incorporated herein by reference in its entirety.

Technical Field

The present disclosure relates generally to compounds, compositions and methods of preparation and use of the compounds and compositions for treating cancer or autoimmune diseases.

Background

BCL6 (B cell lymphoma 6) is a member of the BTB/POZ-zinc finger family, which contains an N-terminal BTB/POZ domain and a zinc finger at the C-terminal end. BCL6, a transcription factor for follicular helper T (Tfh) cells, is required for Germinal Center (GC) formation of naive B cells and thus for antibody affinity maturation. BCL6 was originally discovered as an oncogene in diffuse large B-cell lymphoma (DLBCL) and its role was associated with many types of diseases including B-acute lymphoblastic leukemia, chronic myeloid leukemia, breast cancer, and non-small cell lung cancer (NSCLC) (Cardenas et al, CLIN CANCER RES 2017, 23, 885-893). The N-terminal BTB/POZ domain binds to and recruits co-repressor molecules such as SMRT, NCOR1 and BCOR to form class I and class II histone deacetylase complexes, and the C-terminal zinc finger binds to specific DNA recognition sequences (Yang et al, cell Dev. Biol. 2019, 7, 272). BCL6 reduces RNA expression of its target (including several key tumor suppressors) when bound to its target gene and form a complex. Overexpression of BCL6, commonly found in malignancies such as non-hodgkin lymphomas (NHL), results in ectopic inhibition of cell cycle and DNA repair checkpoint proteins, leading to unrestricted cell proliferation and tumorigenesis.

GC reactions are known to cause increased production of pathogenic autoantibodies leading to several diseases, indicating potential therapeutic applicability of methods of inhibiting or degrading BCL 6. Structural characterization of the co-crystal structure of the BCL6 BTB/POZ domain and the co-repressor suggests that binding occurs at the lateral groove formed by the interface between BCL6 BTB/POZ homodimers (Melnick et al mol, cell biol 2002, 22, 1804-1818; ghetu et al mol, cell 2008, 29, 384-391). Since then, specific ligands that bind to this site have been studied in order to exploit the binding affinity to the lateral groove to make BCL6 a patentable target.

Protein degradation is a highly regulated and vital process to maintain cellular homeostasis. Selective identification and removal of damaged, misfolded or excess proteins is achieved by the ubiquitin-proteasome pathway (UPP). UPP is central to regulating almost all cellular processes. Ubiquitination of proteins is accomplished by an E3 ubiquitin ligase that binds to the protein and adds ubiquitin molecules to the protein, thereby labeling the protein for proteasome degradation.

There has been great interest in utilizing UPP for therapeutic use (Zhou et al mol. Cell 2000, 6, 751-756). One promising therapy uses proteolytically targeted chimeras (commonly referred to as PROTAC) to achieve removal of unwanted proteins by protein degradation (SCHEEPSTRA et al, comp. Struct. Biotech. J. 2019, 17, 160-176). PROTAC are ligand-directed degradants that bind together the E3 ligase and the target protein to be degraded. These divalent molecules typically consist of an E3 ligase ligand linked by a linker moiety to a small molecule that binds the target protein. PROTAC localize the E3 ligase at the appropriate distance and orientation from the target protein, allowing the latter to ubiquitinate. The ubiquitinated target protein is then recognized by the proteasome, where it is degraded.

Thus, in one aspect, provided herein are compounds that target BCL6 for degradation.

Disclosure of Invention

In certain embodiments, described herein are compounds and compositions thereof for modulating BCL 6. In various embodiments, the compounds and compositions thereof are useful for treating cancer.

Embodiments of the present invention may be understood more fully by reference to the detailed description and examples, which are intended to illustrate non-limiting embodiments.

Drawings

FIG. 1A schematic representation of BMS-986458 as a mechanism of BCL 6-LDD. Compound-induced proximity to CRBN E3 ligase promotes ubiquitination of BCL6, resulting in degradation by the 26S proteasome. In DLBCL, loss of BCL6 leads to cell cycle arrest, loss of DNA damage repair, and eventual cell death by apoptosis.

FIG. 2 BCL6 protein levels assessed by MSD-ECL assays at 2h and 4h in (A) OCI-LY-1 and (B) WSU-DLCL-2, respectively, after treatment with BMS-986458. The relative BCL6 protein levels (C) and 5 day proliferation response (D) of the two diastereomers BMT-648592 (filled circles) and BMT-648599 (filled diamonds) relative to BMS-986458 (1:1 mixture) (filled squares) were independently evaluated in OCI-LY-1. The OCI-LY-1 cells treated with 10 nM BMS-986458 achieved complete degradation of BCL6 within 2h (F) and maintained greater than 90% BCL6 degradation for 5 days when dosed at 10 nM (G), as demonstrated by western blotting. No 5-day antiproliferative response (H) to BMS-986458 observed in the OCI-LY-1 parental control was observed in OCI-LY-1 CRBN-/-. Concentrations are listed in μm. Abbreviations DC50, concentration at half maximum degradation, DMSO, dimethyl sulfoxide, POC, percent of DMSO control.

FIG. 3 mass spectrometry proteomic analysis of OCI-LY-1 cells treated with BMS-986458. Volcanic plot exhibits a high substrate specificity of compound-induced degradation upon treatment of 4h (a). BCL6 degradation was maintained at 24 h, however the level of downstream functional effectors was also affected (B). Historical neo-substrates (IKZF 1, IKZF2, IKZF3, ZFP91, CSNK1A1, GSPT 1) associated with CRBN-based degradation mechanisms are highlighted in blue. A statistical robustness threshold was established, with FDR p value < 0.05 highlighted in yellow and red squares. A meaningful cutoff for biological effects is established at log fold change = +/-1, vertical red dashed line. Concentrations are listed in μm. Abbreviation FDR, error discovery Rate.

Fig. 4 acute and delayed anti-proliferative responses following BMS-986458 treatment. AUC values (. Mu.M.h) describe antiproliferative responses to BMS-986458 over 5 days (diagonal filled) and 10 days (filled) of treatment in the equivalent dose range (0.015-100 nM). 10-day DRCs were regulated by proliferation factors obtained after 5-day treatment on DMSO-only controls. Activity was assessed using CTG luminescent cell viability assay. GCB COO is indicated by ().

FIG. 5 shows the apoptotic and antiproliferative responses to BMS-986458 in acute and delayed-response cell lines. In six DLBCL cell line models representing both acute and delayed response classifications, viability (triangle data points, solid line regression; left axis) and apoptosis (circle data points, dashed line regression; right axis) after exposure to BMS-896458 dose ranges up to 40nM for 5 days. Flow cytometry was used to assess apoptotic activity. Abbreviations PoC, percent of control, FC, flow cytometry, AUC, area under concentration-time curve.

FIG. 6 correlation of BMS-986458 antiproliferative response with BCL6 expression levels. Ordering of 10 day AUC values from DLBCL group cell lines in response to BMS-986458 (6A). Ten days AUC < 7 μm.h threshold was used to define the late-onset sensitivity model and the drug-resistance model as indicated by the solid black line. BMS-986458 sensitivity, expressed by the BCL6 baseline protein expression level, was ranked (6B).

FIG. 7 shows the antiproliferative and apoptotic responses of the doxorubicin-resistant DLBCL model to BMS-986458. Viability and apoptosis curves after exposure to serial dilutions of doxorubicin or BMS-986458 for 5 days in three acquired doxorubicin-resistance (dashed line) and corresponding matched parental DLBCL cell lines (solid line) (a). The DLBCL model of acquired doxorubicin resistance and its corresponding matched parental lines responded to western blot protein analysis of 10 nM single dose BMS-986458 over a 72 h treatment period (B). Abbreviation DoxR, doxorubicin resistance.

FIG. 8 plasma/tumor exposure and percent BCL6 degradation in OCI-LY-1 xenografts following BID x5 or BID x3 administration of 30 mg/kg BMS-986458. Female CB.17-SCID mice with approximately 500 mm ³ OCI-LY-1 tumors were dosed twice daily with BMS-986458 at 30 mg/kg for five consecutive days (BID×5, AP 9506) or three consecutive days (BID×3, AP 9721) (n=4/time point). Plasma and tumor samples were harvested at 6, 10 and 14 h post-final dose to determine BMS-986458 concentration. Tumors were treated to detect BCL6 levels with western blots (upper image). Plasma (blue line) and tumor (red line) concentrations (left axis) of BMS-986458 are presented in the graphs of the lower graph. The green bars represent the percent BCL6 degradation in BMS-986458 treated tumors compared to vehicle. The percentage BCL6 degradation was calculated relative to vehicle control (0% BCL6 degradation). All treatment groups were statistically analyzed using one-way anova with Dunnett multiple comparison test (Dunnett's multiple comparisons test) and the data shown were compared to vehicle control groups. The results are presented as average values, where the error bars represent the standard error of the average values. Abbreviations BCL6, B cell lymphoma 6;h hours, BIDx5 twice daily for 5 consecutive days, BIDx3 twice daily for 3 consecutive days, p < 0.0001.

FIG. 9 anti-tumor activity of BMS-986458 (study AP 9776) with twice daily dosing (BID) in OCI-LY-1 xenograft model for 21 consecutive days. Female CB.17-SCID mice were inoculated into the right flank with 5X 10 ⁶ OCI-LY-1 tumor cells in 1:1 matrigel. At the beginning of treatment, mice were randomly assigned to each treatment group (n=9/group). Test article treatment began on day 19 with an average tumor volume of approximately 196 mm ³. The tumor volume reduction was calculated as the center difference in average tumor volume between BMS-986458 treated mice and vehicle treated mice. Day 37, the final tumor measurement day. The results are presented as average values, with error bars representing SEM. All treatment groups were statistically analyzed using a two-way anova with Bonferroni multiple comparison test (Bonferroni's multiple comparisons test) and the data shown were compared to vehicle control groups. Abbreviation BID, twice daily administration, vehicle, 10% Captisol, pH 3; × p < 0.0001 in 25 mM citrate buffer.

Fig. 10 BCL6 degradation in OCI-LY-1 tumors treated with twice daily dosing BMS-986458 for 21 consecutive days collected at 6 h (a), 10 h (B) and 14 h (C) after the final dose. Starting on day 19, the average tumor volume was about 196 mm ³. Three days after final tumor measurement, each BMS-986458 treatment group (n=9) was divided into three cohorts (3 mice per cohort) on day 40 for PD and PK sampling. Tumors were collected 6, 10 and 14 h after the final dose and split into two halves for PD and PK analysis. For PD analysis, tumors were processed into lysates and analyzed for BCL6 protein degradation by western blotting (upper panel, blot image). Plasma and tumor BMS-986458 concentrations and BCL6 degradation are depicted in the lower panel (graph). The results are presented as average integrated intensity values, where the error bars represent the standard error of the average. All treatment groups were statistically analyzed using one-way analysis of variance (ANOVA) with danniter multiple comparison test, and the data shown were compared to vehicle control groups. Abbreviation BID, twice daily administration, vehicle 10% Captisol in 25 mM citrate buffer, pH 3, & p < 0.0001 female CB.17-SCID mice were inoculated into the right flank with 5X 10 ⁶ OCI-LY-1 tumor cells in 1:1 matrigel.

FIG. 11 BMS-986458 treatment induces loss of follicular helper T cell identity and perturbation of cytokine secretion. Phenotypic responses to BMS-986458 were evaluated in iTfh isolated from PBMC from three independent healthy donors. A shows the treatment schedule followed by naive cd4+cd45ra+ T cell induction iTfh differentiation and the time setting of flow analysis using the gating strategy shown (B). Analysis of iTfh on day 4 showed induction of BCL6 expression, expression of the Tfh surface marker CXCR5/PD1, and secretion of canonical Tfh cytokine IL-21 (C). BMS-986458 dose and time dependent modulation (D-F) of BCL6 protein and Tfh phenotype obtained by loss of CXCR5+PD1+ as determined by flow cytometry. IL-10 and IL-121 secretion was assessed using ECL-MSD on days 4 and 8, respectively, after BMS-986458 treatment (Panel G). Nonlinear regression analysis was performed using GRAPHPAD PRISM v 9.4.0. Significance of donor changes in all assays was assessed using a 2 factor anova with significance = p-value > 0.05 (panel H). Abbreviations ActA, activin a, iTfh, follicular helper T cells, D4, day 4, DMSO, dimethyl sulfoxide, ns are not significant.

FIG. 12 BMS-986458 treatment did not affect isolated neutrophil viability or maturation kinetics. Cd34+ cells derived from HD bone marrow were exposed to BMS-986458 at the indicated concentrations for 14 days, with treatment and cell culture medium refreshed every two days. Neutrophil maturation stage was assessed over time by flow cytometry. Data represent the average percentage of cells in each bone marrow population in three donors. Abbreviations DMSO, dimethyl sulfoxide.

FIG. 13A is a flow chart of the BOIN design in stage 1 of section A1 of example B4.

FIG. 13B is a flow chart of the TITE-BOIN design in section A1, section 2 of example B4.

Detailed Description

Definition of the definition

As used herein, the terms "comprising" and "including" are used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of the stated features or components as referred to, but not excluding the presence or addition of one or more features, or components, or groups thereof. In addition, the terms "comprising" and "including" are intended to include examples encompassed by the term "consisting of. Thus, the term "consisting of" may be used in place of the terms "comprising" and "including" to provide a more specific embodiment of the invention.

The term "consisting of means that a subject has at least 90%, 95%, 97%, 98% or 99% of the stated features or components that make up the subject. In another embodiment, the term "consisting of" excludes any other feature or component from the scope of any subsequent enumeration, except for those features or components not necessary for the technical effect to be achieved.

As used herein, the term "or" should be construed as an inclusive "or" meaning any one or any combination. Accordingly, "A, B or C" means any of "A, B, C, A and B, A and C, B and C, A, B and C". An exception to this definition will occur only when a combination of elements, functions, steps or acts are in some way inherently mutually exclusive.

In the description of the present invention, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer within the recited range and (where appropriate) fractions thereof (such as tenths and hundredths of integers). Furthermore, unless otherwise indicated, any numerical range recited herein in connection with any physical feature (such as a polymer subunit, size, or thickness) should be understood to include any integer within the recited range. As used herein, unless otherwise indicated, the terms "about" and "approximately" mean ± 20%, ±10%, ±5% or ± 1% of the indicated range, value or structure.

"Amino" refers to the-NH ₂ group.

"Cyano" refers to a-CN group.

"Nitro" refers to the-NO ₂ group.

"Oxa" refers to an-O-group.

"Oxo" refers to an =o group.

"Sulfur subunit" refers to the = S group.

"Imino" refers to a = N-H group.

"Oximo" refers to an =n-OH group.

"Hydrazino" refers to the = N-NH ₂ group.

"Alkyl" refers to a straight or branched hydrocarbon chain group consisting of only carbon and hydrogen atoms, free of unsaturation, having one to fifteen carbon atoms (e.g., a C ₁-C₁₅ alkyl group). In certain embodiments, the alkyl group comprises one to thirteen carbon atoms (e.g., a C ₁-C₁₃ alkyl group). In certain embodiments, the alkyl group comprises one to eight carbon atoms (e.g., a C ₁-C₈ alkyl group). In other embodiments, the alkyl group comprises one to five carbon atoms (e.g., a C ₁-C₅ alkyl group). In other embodiments, the alkyl group comprises one to four carbon atoms (e.g., a C ₁-C₄ alkyl group). In other embodiments, the alkyl group comprises one to three carbon atoms (e.g., a C ₁-C₃ alkyl group). in other embodiments, the alkyl group comprises one to two carbon atoms (e.g., a C ₁-C₂ alkyl group). In other embodiments, the alkyl group comprises one carbon atom (e.g., a C ₁ alkyl group). In other embodiments, the alkyl group comprises five to fifteen carbon atoms (e.g., a C ₅-C₁₅ alkyl group). In other embodiments, the alkyl group comprises five to eight carbon atoms (e.g., a C ₅-C₈ alkyl group). In other embodiments, the alkyl group comprises two to five carbon atoms (e.g., a C ₂-C₅ alkyl group). In other embodiments, the alkyl group comprises three to five carbon atoms (e.g., a C ₃-C₅ alkyl group). in other embodiments, the alkyl group is selected from the group consisting of methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). The alkyl group is attached to the remainder of the molecule by a single bond. Unless specifically stated otherwise in the specification, alkyl is optionally substituted with one or more of halo, cyano, nitro, oxo, thioylene (thioxo), imino, oximino, trimethylsilyl 、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)- N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -S (O) _tOR^a wherein t is 1 or 2, -S (O) _tR^a wherein t is 1 or 2, and-S (O) _tN(R^a)₂ wherein t is 1 or 2, wherein each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Alkyl-OH" refers to an unbranched or branched alkyl group as defined above wherein one or more hydrogen atoms are replaced by-OH. For example, "C ₁-C₆ alkyl-OH" refers to C ₁-C₆ alkyl substituted with one or more-OH groups. The alkyl-OH may contain multiple hydroxyl groups attached to the same carbon atom or carbon atoms.

"Alkoxy" refers to a group of formula-O-alkyl bonded via an oxygen atom, wherein alkyl is an alkyl chain as defined above.

"Alkenyl" refers to a straight or branched hydrocarbon chain group consisting of only carbon and hydrogen atoms, containing at least one carbon-carbon double bond and having from twenty to twelve carbon atoms. In certain embodiments, alkenyl groups comprise two to eight carbon atoms. In other embodiments, alkenyl groups comprise two to four carbon atoms. Alkenyl is attached to the remainder of the molecule by a single bond, for example, vinyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1, 4-dienyl, and the like. Unless specifically stated otherwise in the specification, alkenyl is optionally substituted with one or more of halo, cyano, nitro, oxo, thiomethylene, imino, oxime, trimethylsilyl 、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)- N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -S (O) _tOR^a wherein t is 1 or 2, -S (O) _tR^a wherein t is 1 or 2, and-S (O) _tN(R^a)₂ wherein t is 1 or 2, wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

"Alkynyl" refers to a straight or branched hydrocarbon chain radical consisting of only carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, having from twenty to twelve carbon atoms. In certain embodiments, alkynyl groups contain two to eight carbon atoms. In other embodiments, alkynyl groups have two to four carbon atoms. Alkynyl groups are attached to the remainder of the molecule by single bonds, e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless specifically stated otherwise in the specification, alkynyl is optionally substituted with one or more of halo, cyano, nitro, oxo, thiomethylene, imino, oxime, trimethylsilyl 、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)- N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -S (O) _tOR^a wherein t is 1 or 2, -S (O) _tR^a wherein t is 1 or 2, and-S (O) _tN(R^a)₂ wherein t is 1 or 2, wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, methoxy, hydroxy, or trifluoromethyl).

"Alkylene" or "alkylene chain" refers to a straight or linear divalent hydrocarbon chain linking the remainder of the molecule to a group, consisting of only carbon and hydrogen, free of unsaturation and having one to twelve carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the remainder of the molecule via a single bond and to the group via a single bond. The attachment of the alkylene chain to the remainder of the molecule and to the group may be via one carbon in the alkylene chain or via any two carbons in the chain. In certain embodiments, the alkylene group comprises one to eight carbon atoms (e.g., a C ₁-C₈ alkylene group). In other embodiments, the alkylene group comprises one to five carbon atoms (e.g., a C ₁-C₅ alkylene group). In other embodiments, the alkylene group comprises one to four carbon atoms (e.g., a C ₁-C₄ alkylene group). In other embodiments, the alkylene group comprises one to three carbon atoms (e.g., a C ₁-C₃ alkylene group). In other embodiments, the alkylene group comprises one to two carbon atoms (e.g., a C ₁-C₂ alkylene group). In other embodiments, the alkylene group comprises one carbon atom (e.g., a C ₁ alkylene group). In other embodiments, the alkylene group comprises five to eight carbon atoms (e.g., a C ₅-C₈ alkylene group). in other embodiments, the alkylene group comprises two to five carbon atoms (e.g., a C ₂-C₅ alkylene group). In other embodiments, the alkylene group comprises three to five carbon atoms (e.g., a C ₃-C₅ alkylene group). Unless specifically stated otherwise in the specification, the alkylene chain is optionally substituted with one or more of halo, cyano, nitro, oxo, thioylene, imino, oxime, trimethylsilyl 、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)- N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -S (O) _tOR^a wherein t is 1 or 2, -S (O) _tR^a wherein t is 1 or 2, and-S (O) _tN(R^a)₂ wherein t is 1 or 2, wherein each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Aryl" refers to a group derived from an aromatic mono-or polycyclic hydrocarbon ring system by removal of a hydrogen atom from a ring carbon atom. An aromatic mono-or polycyclic hydrocarbon ring system contains only hydrogen and carbon from five to eighteen carbon atoms, wherein at least one ring of the ring system is fully unsaturated, i.e. it contains a cyclic delocalized (4n+2) pi-electron system according to the shock theory (Hu ckel theory). Ring systems from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin, and naphthalene. Unless specifically stated otherwise in the specification, the term "aryl" or the prefix "aryl" (such as in "aralkyl") is intended to include aryl optionally substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl 、-R^b-OR^a、-R^b-OC(O)-R^a、-R^b-OC(O)-OR^a、-R^b-OC(O)-N(R^a)₂、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -R ^b-S(O)_tR^a (where t is 1 or 2), -R ^b-S(O)_tOR^a (where t is 1 or 2), and-R ^b-S(O)_tN(R^a)₂ (where t is 1 or 2), where each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), Methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a linear or branched alkylene or alkenylene chain, and R ^c is a linear or branched alkylene or alkenylene chain, and wherein each of the above substituents is unsubstituted unless otherwise indicated.

"Aralkyl" refers to a group of the formula-R ^c aryl, wherein R ^c is an alkylene chain as defined above, e.g., methylene, ethylene, and the like. The alkylene chain portion of the aralkyl group is optionally substituted as described above for the alkylene chain. The aryl portion of the aralkyl is optionally substituted as described above for aryl.

"Aralkenyl" refers to a group of the formula-R ^d -aryl, wherein R ^d is an alkenylene chain as defined above. The aryl moiety of the arylalkenyl is optionally substituted as described above for aryl. The alkenylene moiety of the aralkenyl is optionally substituted as defined above for alkenylene.

"Aralkynyl" refers to a group of the formula-R ^e -aryl, wherein R ^e is an alkynylene chain as defined above. The aryl portion of the arylalkynyl group is optionally substituted as described above for the aryl group. The alkynylene chain portion of the aralkynyl group is optionally substituted as defined above for the alkynylene chain.

"Carbocyclyl" refers to a stable, non-aromatic, monocyclic or multicyclic hydrocarbon group consisting of only carbon and hydrogen atoms, said group comprising a fused or bridged ring system having from three to fifteen carbon atoms. In certain embodiments, carbocyclyl groups contain three to ten carbon atoms. In other embodiments, carbocyclyl groups contain five to seven carbon atoms. The carbocyclyl is attached to the remainder of the molecule by a single bond. Carbocyclyl groups may be saturated (i.e., contain only a single c—c bond) or unsaturated (i.e., contain one or more double or triple bonds). Fully saturated carbocyclyl is also referred to as "carbocyclyl". Examples of monocyclic carbocyclyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Unsaturated carbocyclyl is also known as "cycloalkenyl". Examples of monocyclic cycloalkenyl groups include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl groups include, for example, adamantyl, norbornyl (i.e., bicyclo [2.2.1] heptanyl), norbornyl, decalinyl, 7-dimethylbicyclo [2.2.1] heptanyl, and the like. Unless specifically indicated otherwise in the specification, the term "carbocyclyl" is intended to include carbocyclyl optionally substituted with one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioylene, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl 、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)-N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -S (O) _tOR^a (where t is 1 or 2), -S (O) _tR^a (where t is 1 or 2), and-S (O) _tN(R^a)₂ (where t is 1 or 2), where each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), Methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each Rb is independently a direct bond or a linear or branched alkylene or alkenylene chain, and R ^c is a linear or branched alkylene or alkenylene chain, and wherein each of the above substituents is unsubstituted unless otherwise indicated.

"Carbocyclylalkyl" refers to a group of formula-R ^c -carbocyclyl, wherein R ^c is an alkylene chain as defined above. The alkylene chain and carbocyclyl are optionally substituted as defined above.

"Carbonyl" refers to a group of formula-C (O) R ¹⁰R²⁰, wherein R ¹⁰ and R ²⁰ are independently selected from-OH, halo, cyano, nitro, oxo, thiomethylene, imino, oxime, trimethylsilyl 、-R^a、-OR^a、-SR^a、-OC(O)-R^a、-N(R^a)₂、-C(O)R^a、-C(O)OR^a、-C(O)N(R^a)₂、-N(R^a)C(O)OR^a、-OC(O)- N(R^a)₂、-N(R^a)C(O)R^a、-N(R^a)S(O)_tR^a（, wherein t is 1 or 2), -S (O) _tOR^a (wherein t is 1 or 2), -S (O) _tR^a (wherein t is 1 or 2), and-S (O) _tN(R^a)₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, methoxy, or trifluoromethyl).

"Halo" or "halogen" refers to a bromo, chloro, fluoro or iodo substituent.

"Haloalkyl" refers to an alkyl group as defined above substituted with one or more halo groups as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2-trifluoroethyl, 1, 2-difluoroethyl, 3-bromo-2-fluoropropyl, 1, 2-dibromoethyl, and the like. In some embodiments, haloalkyl has one to six carbon atoms and is substituted with one or more halo groups (C ₁-C₆ haloalkyl), or the haloalkyl has one to five carbon atoms and is substituted with one or more halo groups (C ₁-C₅ haloalkyl), or the haloalkyl has one to three carbon atoms and is substituted with one or more halo groups (C ₁-C₃ haloalkyl). The halo groups may all be the same or the halo groups may be different. Unless otherwise specifically indicated, haloalkyl is optionally substituted.

"Fluoroalkyl" refers to an alkyl group as defined above substituted with one or more fluoro groups as defined above, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, 2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. The alkyl portion of the fluoroalkyl group may be optionally substituted as defined above for the alkyl group.

"Heterocyclyl" refers to a stable 3-to 18-membered non-aromatic ring group containing from twenty-two carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. Unless specifically stated otherwise in the specification, a heterocyclyl is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system that may include a fused or bridged ring system. The heteroatoms in the heterocyclyl may optionally be oxidized. One or more nitrogen atoms (if present) are optionally quaternized. The heterocyclyl groups are partially or fully saturated. The heterocyclyl may be attached to the remainder of the molecule via any atom of one or more rings. Examples of such heterocyclic groups include, but are not limited to, dioxanyl, thienyl [1,3] dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, and 1, 1-dioxothiomorpholinyl. Unless specifically stated otherwise in the specification, the term "heterocyclyl" is intended to include heterocyclyl as defined above, optionally substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioylene, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl 、-R^b-OR^a、-R^b-OC(O)-R^a、-R^b-OC(O)-OR^a、-R^b-OC(O)-N(R^a)₂、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -R ^b-S(O)_tR^a (where t is 1 or 2), -R ^b-S(O)_tOR^a (where t is 1 or 2), and-R ^b-S(O)_tN(R^a)₂ (where t is 1 or 2), where each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), Methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a linear or branched alkylene or alkenylene chain, and R ^c is a linear or branched alkylene or alkenylene chain, and wherein each of the above substituents is unsubstituted unless otherwise indicated.

"N-heterocyclyl" or "N-attached heterocyclyl" refers to a heterocyclyl as defined above which contains at least one nitrogen and wherein the point of attachment of the heterocyclyl to the remainder of the molecule is via a nitrogen atom in the heterocyclyl. The N-heterocyclyl is optionally substituted as described above for the heterocyclyl. Examples of such N-heterocyclyl groups include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

"C-heterocyclyl" or "C-attached heterocyclyl" refers to a heterocyclyl as defined above which contains at least one heteroatom and in which the point of attachment of the heterocyclyl to the remainder of the molecule is via a carbon atom in the heterocyclyl. The C-heterocyclyl is optionally substituted as described above for the heterocyclyl. Examples of such C-heterocyclyl groups include, but are not limited to, 2-morpholinyl, 2-or 3-or 4-piperidinyl, 2-piperazinyl, 2-or 3-pyrrolidinyl, and the like.

"Heterocyclylalkyl" refers to a group of the formula-R ^c -heterocyclyl, wherein R ^c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl at a nitrogen atom. The alkylene chain of the heterocyclylalkyl is optionally substituted as defined above for the alkylene chain. The heterocyclyl portion of the heterocyclylalkyl is optionally substituted as defined above for heterocyclyl.

"Heterocyclylalkoxy" refers to a group bonded via an oxygen atom of a heterocyclic group of formula-O-R ^c, wherein R ^c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl at a nitrogen atom. The alkylene chain of the heterocyclylalkoxy group is optionally substituted as defined above for the alkylene chain. The heterocyclyl portion of the heterocyclylalkoxy group is optionally substituted as defined above for the heterocyclyl.

"Heteroaryl" refers to groups derived from 3 to 18 membered aromatic ring groups containing from two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, heteroaryl groups may be monocyclic, bicyclic, tricyclic or tetracyclic ring systems, wherein at least one ring of the ring system is fully unsaturated, i.e., it contains a cyclic delocalized (4n+2) pi-electron system according to the shock theory. Heteroaryl groups include fused or bridged ring systems. One or more heteroatoms in the heteroaryl group are optionally oxidized. One or more nitrogen atoms (if present) are optionally quaternized. Heteroaryl groups may be attached to the remainder of the molecule via any atom of one or more rings. Examples of heteroaryl groups include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1, 3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo [ d ] thiazolyl, benzothiadiazolyl, benzo [ b ] [1,4] dioxaheptenyl, benzo [ b ] [1,4] oxazinyl, 1, 4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxanyl, benzopyranyl, benzopyronyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothienyl) (benzothienyl (benzothiophenyl)) Benzo [3,2-d ] pyrimidinyl, benzotriazole, benzo [4,6] imidazo [1,2-a ] pyridinyl, carbazolyl, cinnolinyl, cyclopenta [ d ] pyrimidinyl, 6, 7-dihydro-5H-cyclopenta [4,5] thieno [2,3-d ] pyrimidinyl, 5, 6-dihydrobenzo [ H ] quinazolinyl, 5, 6-dihydrobenzo [ H ] cinnolinyl, 6, 7-dihydro-5H-benzo [6,7] cyclohepta [1,2-c ] pyridazinyl, dibenzofuranyl, dibenzothienyl, furanyl, furanonyl, furo [3,2-c ] pyridinyl, 5,6,7,8,9, 10-hexahydrocyclooctatetraene [ d ] pyrimidinyl, 5,6,7,8,9, 10-hexahydrocyclooctatetraene [ d ] pyridazinyl, 5,6,7,8,9, 10-hexahydrocyclooctatetraene [ d ] pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolinyl, indolizinyl, isoxazolyl, 5, 8-methano-5, 6,7, 8-tetrahydroquinazolinyl, naphthyridinyl, 1, 6-naphthyridinyl, oxadiazolyl, 2-oxo-azepinyl, oxazolyl, oxiranyl, 5, 6a,7,8,9,10 a-octahydrobenzo [ H ] quinazolinyl, 1-phenyl-1H-pyrrolyl, Phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo [3,4-d ] pyrimidinyl, pyridinyl, pyrido [3,4-d ] pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7, 8-tetrahydroquinazolinyl, 5,6,7, 8-tetrahydrobenzo [4,5] thieno [2,3-d ] pyrimidinyl, 6,7,8, 9-tetrahydro-5H-cyclohepto [4,5] thieno [2,3-d ] pyrimidinyl, 5,6,7, 8-tetrahydropyrido [4,5-c ] pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno [2,3-d ] pyrimidinyl, thieno [3,2-d ] pyrimidinyl, thieno [2,3-c ] pyridinyl, and thienyl (thiophenyl) (i.e., thienyl (thienyl)). unless specifically stated otherwise in the specification, the term "heteroaryl" is intended to include heteroaryl as defined above, optionally substituted with one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioylidene, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, Optionally substituted heteroarylalkyl 、-R^b-OR^a、-R^b-OC(O)-R^a、-R^b-OC(O)-OR^a、-R^b-OC(O)-N(R^a)₂、-R^b-N(R^a)₂、-R^b-C(O)R^a、-R^b-C(O)OR^a、-R^b-C(O)N(R^a)₂、-R^b-O-R^c-C(O)N(R^a)₂、-R^b-N(R^a)C(O)OR^a、-R^b-N(R^a)C(O)R^a、-R^b-N(R^a)S(O)_tR^a（ wherein t is 1 or 2), -R ^b-S(O)_tR^a (wherein t is 1 or 2), -R ^b-S(O)_tOR^a (wherein t is 1 or 2), and-R ^b-S(O)_tN(R^a)₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, Alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), Methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a linear or branched alkylene or alkenylene chain, and R ^c is a linear or branched alkylene or alkenylene chain, and wherein each of the above substituents is unsubstituted unless otherwise indicated.

"N-heteroaryl" refers to a heteroaryl group as defined above, which contains at least one nitrogen, and wherein the point of attachment of the heteroaryl group to the separate remainder of the molecule is via a nitrogen atom in the heteroaryl group. The N-heteroaryl is optionally substituted as described above for heteroaryl.

"C-heteroaryl" refers to a heteroaryl group as defined above, and wherein the point of attachment of the heteroaryl group to the remainder of the molecule is via a carbon atom in the heteroaryl group. The C-heteroaryl is optionally substituted as described above for the heterocyclyl.

"Heteroarylalkyl" refers to a group of the formula-R ^c -heteroaryl, wherein R ^c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl at a nitrogen atom. The alkylene chain of the heteroarylalkyl is optionally substituted as defined above for the alkylene chain. The heteroaryl portion of the heteroarylalkyl is optionally substituted as defined above for heteroaryl.

"Heteroarylalkoxy" refers to a group bonded via an oxygen atom of a heteroaryl of formula-O-R ^c, wherein R ^c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl at a nitrogen atom. The alkylene chain of the heteroarylalkoxy group is optionally substituted as defined above for the alkylene chain. The heteroaryl portion of the heteroarylalkoxy is optionally substituted as defined above for heteroaryl.

Embodiments of the present disclosure are intended to encompass pharmaceutically acceptable salts, tautomers, isotopologues and stereoisomers of the compounds provided herein, such as compounds of formula (I).

As used herein, the term "one or more pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases (including inorganic acids and bases and organic acids and bases). Suitable pharmaceutically acceptable base addition salts of the compounds of formula (I) include, but are not limited to, metal salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from lysine, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl-glucamine) and procaine. Suitable non-toxic acids include, but are not limited to, inorganic and organic acids such as acetic acid, alginic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethenesulfonic acid, formic acid, fumaric acid, furancarboxylic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, propionic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. Specific non-toxic acids include hydrochloric acid, hydrobromic acid, maleic acid, phosphoric acid, sulfuric acid, and methanesulfonic acid. Thus, examples of specific salts include hydrochloride, formate and mesylate salts. Other are well known in the art, see, for example, remington's Pharmaceutical Sciences, 18 th edition, mack Publishing, easton Pa (1990) or Remington: THE SCIENCE AND PRACTICE of Pharmacy, 19 th edition, mack Publishing, iston, pa (1995).

As used herein and unless otherwise indicated, the term "stereoisomer" or "stereomerically pure" means one stereoisomer of a particular compound that is substantially free of the other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of the compound. A stereomerically pure compound having two chiral centers will be substantially free of the other diastereomers of the compound. Typical stereoisomerically pure compounds comprise more than about 80% by weight of one stereoisomer of a compound and less than about 20% by weight of the other stereoisomers of a compound, more than about 90% by weight of one stereoisomer of a compound and less than about 10% by weight of the other stereoisomers of a compound, more than about 95% by weight of one stereoisomer of a compound and less than about 5% by weight of the other stereoisomers of a compound, or more than about 97% by weight of one stereoisomer of a non-compound and less than about 3% by weight of the other stereoisomers of a compound. The compounds disclosed herein may have chiral centers and may exist as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms are included in the embodiments disclosed herein, including mixtures thereof.

Embodiments disclosed herein encompass the use of stereoisomerically pure forms of the compounds disclosed herein as well as the use of mixtures of those forms. For example, mixtures comprising equal or unequal amounts of enantiomers of a particular compound may be used in the methods and compositions disclosed herein. These isomers may be asymmetrically synthesized or resolved using standard techniques such as chiral columns or chiral resolving agents. See, e.g., jacques, J., et al Enantiomers, RACEMATES and solutions (Wiley-Interscience, N.Y., 1981), wilen, S.H., et al Tetrahedron 33:2725 (1977), eliel, E.L., stereochemistry of Carbon Compounds (McGraw-Hill, N.Y., 1962), wilen, S.H., tables of Resolving AGENTS AND Optical Resolutions, page 268 (E.L. Eliel, eds., univ. Of note DAME PRESS, santa Claus , 1972);Todd, M., Separation Of Enantiomers : Synthetic Methods (Wiley-VCH Verlag GmbH & Co. KGaA, Germany Wei Yinhai, indian, et al , 2014);Toda, F., Enantiomer Separation: Fundamentals and Practical Methods (Springer Science & Business Media, 2007);Subramanian, G. Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008);Ahuja, S., Chiral Separation Methods for Pharmaceutical and Biotechnological Products (John Wiley & Sons, 2011).

"Tautomer" refers to the isomeric forms of a compound that are in equilibrium with each other. The concentration of the isomeric forms will depend on the environment in which the compound is found and may vary depending on, for example, whether the compound is solid or in an organic or aqueous solution. For example, in aqueous solutions, pyrazoles can exhibit the following isomeric forms, which are referred to as tautomers of each other:

。

As will be readily appreciated by those skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism, and all tautomers of the compounds of formula (I) are within the scope of the disclosure.

Polymers or similar indefinite structures resulting from definition of substituents with infinitely additional further substituents (e.g., substituted aryl groups having substituted alkyl groups that are themselves substituted with substituted aryl groups that are further substituted with substituted heteroalkyl groups, etc.) are not intended to be included herein. Similarly, the above definition is not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluorine or heteroaryl having two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to the skilled artisan.

It should also be noted that the compounds disclosed herein may contain non-natural proportions of atomic isotopes at one or more atoms. For example, the compounds may be radiolabeled with a radioisotope such as, for example, tritium (³ H), iodine-125 (¹²⁵ I), sulfur-35 (³⁵ S), or carbon-14 (¹⁴ C), or may be isotopically enriched such as with deuterium (² H), carbon-13 (¹³ C), or nitrogen-15 (¹⁵ N). As used herein, "isotopologue" is an isotopically enriched compound. The term "isotopically enriched" refers to an atom having an isotopic composition different from the natural isotopic composition of the atom. "isotopically enriched" may also refer to compounds containing at least one atom whose isotopic composition differs from the natural isotopic composition of the atom. The term "isotopic composition" refers to the amount of each isotope present for a given atom. Radiolabeled and isotopically enriched compounds are useful as therapeutic agents, for example, cancer therapeutic agents, research reagents (e.g., binding assay reagents), and diagnostic reagents (e.g., in vivo imaging agents). All isotopic variations of the compounds as described herein, whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, isotopologues of the compounds disclosed herein are provided, e.g., isotopologues are deuterium, carbon-13, and/or nitrogen-15 enriched compounds. As used herein, "deuterated" means a compound in which at least one hydrogen (H) has been replaced with deuterium (indicated by D or ² H), i.e., the compound is enriched in deuterium at least one position.

The disclosure also includes "deuterated analogs" of the compounds described herein wherein 1 to n hydrogens attached to a carbon atom are replaced with deuterium, where n is the number of hydrogens in the molecule. When multiple deuterium atoms are present in a compound, the deuterium atoms can be on the same portion of the molecule (e.g., on a single alkyl group or a single ring) or on different portions of the molecule (e.g., on separate alkyl groups or separate rings). Such compounds may exhibit increased metabolic resistance and, thus, may be useful for increasing the half-life of any compound when administered to a mammal (particularly a human). See, e.g., ,Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci. 5(12):524-527 (1984). such compounds are synthesized by means well known in the art, e.g., by employing starting materials in which one or more hydrogens have been replaced with deuterium.

It is to be understood that each compound disclosed herein may be provided in the form of any pharmaceutically acceptable salt discussed herein, independent of stereoisomers or isotopic composition. Likewise, it is understood that the isotopic composition may vary independently of the stereoisomeric composition of each compound mentioned herein. Furthermore, the isotopic composition, although limited to those elements present in the corresponding compounds disclosed herein or salts thereof, can additionally vary independently of the selection of a pharmaceutically acceptable salt of the corresponding compound.

It should be noted that if there is a difference between the depicted structure and the name of the structure, the depicted structure will be given more weight.

As used herein, "treating" or "treatment" means the complete or partial alleviation of a disorder, disease or condition, or one or more symptoms associated with a disorder, disease or condition, or the slowing or stopping of further progression or worsening of such symptoms, or the alleviation or elimination of one or more causes of the disorder, disease or condition itself. In one embodiment, the disorder is cancer or a symptom thereof as described herein.

As used herein, "preventing" means a method of delaying and/or excluding all or part of the onset, recurrence or spread of a disorder, disease or condition, preventing a subject from acquiring a disorder, disease or condition, or reducing the risk of a subject acquiring a disorder, disease or condition. In one embodiment, the disorder is cancer or a symptom thereof as described herein.

The term "effective amount" in relation to a compound disclosed herein means an amount capable of treating or preventing a disorder, disease or condition disclosed herein, or a symptom thereof.

As used herein, the term "subject" or "patient" includes animals, including but not limited to animals such as cows, monkeys, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, in one embodiment mammals, and in another embodiment humans. In one embodiment, the subject is a human suffering from or at risk of suffering from a BCL 6-mediated disease or symptoms thereof.

Although various features of the invention may be described in the context of a single embodiment, the features can also be provided separately or in any suitable combination. Conversely, although the invention may be described herein in the context of separate embodiments for clarity, the invention may also be implemented in a single embodiment.

Compounds of formula (I)

In one aspect, provided herein is a compound of formula (IA):

(IA)

or a pharmaceutically acceptable salt thereof, wherein:

X ¹ and X ² are each independently N or CH, provided that at least one of X ¹ and X ² is N;

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

R ^2a and R ^2b are each independently H, C ₁-C₆ alkyl, -O (C ₁-C₆ alkyl), -OH, halo, C ₁-C₆ haloalkyl or C ₁-C₆ alkyl-OH,

Or R ^2a and R ^2b together with the carbon atom to which they are attached form a spiro C ₃-C₅ cycloalkyl,

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

Or R ^4a and R ^4b together with the carbon atom to which they are attached form a spiro C ₃-C₅ cycloalkyl group;

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

R ¹¹ is H, C ₁-C₆ alkyl, - (C ₁-C₆ alkylene) - (5-to 6-membered heterocyclyl), - (C ₁-C₆ alkylene) -O (C ₁-C₆ alkyl), C ₁-C₆ haloalkyl, C ₁-C₆ alkyl-OH or- (C ₁-C₆ alkylene) -NH (C ₁-C₆ alkyl),

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

Wherein one or more hydrogen atoms in the compound are optionally replaced with deuterium.

In a further aspect, provided herein is a compound of formula (I):

(I)

or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

R ^2a and R ^2b are each independently H, C ₁-C₆ alkyl, -OH, halo or C ₁-C₆ alkyl-OH,

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

In some embodiments, X ¹ is CH and X ² is N. In some embodiments, X ¹ is N and X ² is CH. In some embodiments, both X ¹ and X ² are N.

In some embodiments, R ^1a is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^1a is H, halo, or C ₁-C₃ alkyl. In some embodiments, R ^1a is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^1a is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^1a is H.

In some embodiments, R ^1a is halo. In some embodiments, R ^1a is F, cl, br, or I. In some embodiments, R ^1a is F.

In some embodiments, R ^1a is C ₁-C₆ alkyl. In some embodiments, R ^1a is C ₁-C₃ alkyl. In some embodiments, R ^1a is methyl, ethyl, or propyl. In some embodiments, R ^1a is methyl.

In some embodiments, R ^1b is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^1b is H, halo, or C ₁-C₃ alkyl. In some embodiments, R ^1b is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^1b is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^1b is H.

In some embodiments, R ^1b is halo. In some embodiments, R ^1b is F, cl, br, or I. In some embodiments, R ^1b is F.

In some embodiments, R ^1a and R ^1b are each independently H, halo, or C ₁-C₃ alkyl. In some embodiments, R ^1a and R ^1b are each independently H, F, cl, br, I, methyl, ethyl, or propyl. In some embodiments, R ^1a and R ^1b are each independently H or-CH ₃.

In some embodiments, R ^1b is C ₁-C₆ alkyl. In some embodiments, R ^1b is C ₁-C₃ alkyl. In some embodiments, R ^1b is methyl, ethyl, or propyl. In some embodiments, R ^1b is methyl.

In some embodiments, R ^2a is H, C ₁-C₆ alkyl, -O (C ₁-C₆ alkyl), -OH, halo, C ₁-C₆ haloalkyl, or C ₁-C₆ alkyl-OH. In some embodiments, R ^2a is H, C ₁-C₃ alkyl, -O (C ₁-C₃ alkyl), -OH, F, cl, br, I, C ₁-C₃ haloalkyl, or C ₁-C₃ alkyl-OH. In some embodiments, R ^2a is H, methyl, ethyl, propyl 、-OCH₃、-OH、F、Cl、Br、I、-CF₃、-CHF₂、-CCl₃、-CH₂OH、-(CH₂CH₂)OH, or- (CH ₂CH₂CH₂) OH.

In some embodiments, R ^2a is H.

In some embodiments, R ^2a is C ₁-C₆ alkyl. In some embodiments, R ^2a is C ₁-C₃ alkyl. In some embodiments, R ^2a is methyl, ethyl, or propyl. In some embodiments, R ^2a is methyl.

In some embodiments, R ^2a is-O (C ₁-C₆ alkyl). In some embodiments, R ^2a is-O (C ₁-C₃ alkyl). In some embodiments, R ^2a is-OCH ₃、-OCH₂CH₃、-OCH₂CH₂CH₃ or-OCH (CH ₃)₂. In some embodiments, R ^2a is-OCH ₃.

In some embodiments, R ^2a is —oh.

In some embodiments, R ^2a is halo. In some embodiments, R ^2a is F, cl, br, or I. In some embodiments, R ^2a is F.

In some embodiments, R ^2a is C ₁-C₆ haloalkyl. In some embodiments, R ^2a is C ₁-C₆ haloalkyl containing 1 to 13 halogen atoms. In some embodiments, R ^2a is C ₁-C₃ haloalkyl. In some embodiments, R ^2a is C ₁-C₃ haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R ^2a is -CF₃、-CHF₂、-CH₂F、-CCl₃、-CHCl₂、-CH₂Cl、-CF₂Cl、-CFCl₂、-CH₂CF₃、-CH₂CHF₂ or-CH ₂CCl₃. In some embodiments, R ^2a is-CF ₃.

In some embodiments, R ^2a is C ₁-C₆ alkyl-OH. In some embodiments, R ^2a is C ₁-C₃ alkyl-OH. In some embodiments, R ^2a is-CH ₂OH、-(CH₂CH₂) OH or- (CH ₂CH₂CH₂) OH. In some embodiments, R ^2a is-CH ₂ OH.

In some embodiments, R ^2b is H, C ₁-C₆ alkyl, -OH, halo, or C ₁-C₆ alkyl-OH. In some embodiments, R ^2b is H, C ₁-C₃ alkyl, -OH, F, cl, br, I, or C ₁-C₃ alkyl-OH. In some embodiments, R ^2b is H, methyl, ethyl, propyl, -OH, F, cl, br, I, -CH ₂OH、-(CH₂CH₂) OH or- (CH ₂CH₂CH₂) OH.

In some embodiments, R ^2b is H.

In some embodiments, R ^2b is C ₁-C₆ alkyl. In some embodiments, R ^2b is C ₁-C₃ alkyl. In some embodiments, R ^2b is methyl, ethyl, or propyl. In some embodiments, R ^2b is methyl.

In some embodiments, R ^2b is-O (C ₁-C₆ alkyl). In some embodiments, R ^2b is-O (C ₁-C₃ alkyl). In some embodiments, R ^2b is-OCH ₃、-OCH₂CH₃、-OCH₂CH₂CH₃ or-OCH (CH ₃)₂. In some embodiments, R ^2b is-OCH ₃.

In some embodiments, R ^2b is —oh.

In some embodiments, R ^2b is halo. In some embodiments, R ^2b is F, cl, br, or I. In some embodiments, R ^2b is F.

In some embodiments, R ^2b is C ₁-C₆ haloalkyl. In some embodiments, R ^2b is C ₁-C₆ haloalkyl containing 1 to 13 halogen atoms. In some embodiments, R ^2b is C ₁-C₃ haloalkyl. In some embodiments, R ^2b is C ₁-C₃ haloalkyl containing 1 to 7 halogen atoms. In some embodiments, R ^2b is -CF₃、-CHF₂、-CH₂F、-CCl₃、-CHCl₂、-CH₂Cl、-CF₂Cl、-CFCl₂、-CH₂CF₃、-CH₂CHF₂ or-CH ₂CCl₃. In some embodiments, R ^2b is-CF ₃.

In some embodiments, R ^2b is C ₁-C₆ alkyl-OH. In some embodiments, R ^2b is C ₁-C₃ alkyl-OH. In some embodiments, R ^2b is-CH ₂OH、-(CH₂CH₂) OH or- (CH ₂CH₂CH₂) OH. In some embodiments, R ^2b is-CH ₂ OH.

In some embodiments, R ^2a and R ^2b together with the carbon atom to which they are attached form a spiro C ₃-C₅ cycloalkyl group. In some embodiments, R ^2a and R ^2b together with the carbon atom to which they are attached form a spiro C ₃-C₄ cycloalkyl group. In some embodiments, R ^2a and R ^2b together with the carbon atom to which they are attached form a spirocyclobutyl.

In some embodiments, R ^2a and R ^2b together form oxo.

In some embodiments, R ^1a and R ^1b are both H, and at least one of R ^2a and R ^2b is not H.

In some embodiments, R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene. In some embodiments, R ^1a and R ^2a together form a bridging ethylene group.

In some embodiments, R ^3a is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^3a is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^3a is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^3a is H.

In some embodiments, R ^3a is halo. In some embodiments, R ^3a is F, cl, br, or I. In some embodiments, R ^3a is F.

In some embodiments, R ^3a is C ₁-C₆ alkyl. In some embodiments, R ^3a is C ₁-C₃ alkyl. In some embodiments, R ^3a is methyl, ethyl, or propyl. In some embodiments, R ^3a is methyl.

In some embodiments, R ^3b is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^3b is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^3b is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^3b is H.

In some embodiments, R ^3b is halo. In some embodiments, R ^3b is F, cl, br, or I. In some embodiments, R ^3b is F.

In some embodiments, R ^3b is C ₁-C₆ alkyl. In some embodiments, R ^3b is C ₁-C₃ alkyl. In some embodiments, R ^3b is methyl, ethyl, or propyl. In some embodiments, R ^3b is methyl.

In some embodiments, R ^4a is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^4a is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^4a is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^4a is H.

In some embodiments, R ^4a is halo. In some embodiments, R ^4a is F, cl, br, or I. In some embodiments, R ^4a is F.

In some embodiments, R ^4a is C ₁-C₆ alkyl. In some embodiments, R ^4a is C ₁-C₃ alkyl. In some embodiments, R ^4a is methyl, ethyl, or propyl. In some embodiments, R ^4a is methyl.

In some embodiments, R ^4b is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^4b is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^4b is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^4b is H.

In some embodiments, R ^4b is halo. In some embodiments, R ^4b is F, cl, br, or I. In some embodiments, R ^4b is F.

In some embodiments, R ^4b is C ₁-C₆ alkyl. In some embodiments, R ^4b is C ₁-C₃ alkyl. In some embodiments, R ^4b is methyl, ethyl, or propyl. In some embodiments, R ^4b is methyl.

In some embodiments, R ^4a and R ^4b together with the carbon atom to which they are attached form a spiro C ₃-C₅ cycloalkyl group. In some embodiments, R ^4a and R ^4b together with the carbon atom to which they are attached form a spiro C ₃-C₄ cycloalkyl group. In some embodiments, R ^4a and R ^4b together with the carbon atom to which they are attached form a spirocyclobutyl.

In some embodiments, R ^3a、R^3b、R^4a and R ^4b are each H.

In some embodiments, at least one of R ^3a and R ^3b is not H, and at least one of R ^4a and R ^4b is not H.

In some embodiments, R ^5a is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^5a is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^5a is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^5a is H.

In some embodiments, R ^5b is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ^5b is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ^5b is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ^5b is H.

In some embodiments, R ^5a and R ^5b are each H.

In some embodiments, at least one of R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a and R ^5b is not H.

In some embodiments, one, two, or three of R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a and R ^5b are not H.

In some embodiments, one or both of R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a and R ^5b is not H.

In some embodiments, w is 0. In some embodiments, w is 1.

In some embodiments of the present invention, in some embodiments,The method comprises the following steps:

。

In some embodiments, x and y are each 0. In some embodiments, x is 0 and y is 1. In some embodiments, x is 1 and y is 0.

In some embodiments, R ⁶ is H or C ₁-C₆ alkyl. In some embodiments, R ⁶ is H or C ₁-C₃ alkyl. In some embodiments, R ⁶ is H, methyl, ethyl, or propyl.

In some embodiments, R ⁶ is H.

In some embodiments, R ⁶ is C ₁-C₆ alkyl. In some embodiments, R ⁶ is C ₁-C₃ alkyl. In some embodiments, R ⁶ is methyl, ethyl, or propyl. In some embodiments, R ⁶ is methyl.

In some embodiments, x is 0, y is 1 and R ⁶ is H.

In some embodiments, x is 0, y is 1 and R ⁶ is C ₁-C₆ alkyl. In some embodiments, x is 0, y is 1 and R ⁶ is C ₁-C₃ alkyl. In some embodiments, x is 0, y is 1 and R ⁶ is methyl.

In some embodiments, R ⁷ is H or C ₁-C₆ alkyl. In some embodiments, R ⁷ is H or C ₁-C₃ alkyl. In some embodiments, R ⁷ is H, methyl, ethyl, or propyl.

In some embodiments, R ⁷ is H.

In some embodiments, R ⁷ is C ₁-C₆ alkyl. In some embodiments, R ⁷ is C ₁-C₃ alkyl. In some embodiments, R ⁷ is methyl, ethyl, or propyl. In some embodiments, R ⁷ is methyl.

In some embodiments, x is 1, y is 0 and R ⁷ is H.

In some embodiments, x is 1, y is 0 and R ⁷ is C ₁-C₆ alkyl. In some embodiments, x is 1, y is 0 and R ⁷ is C ₁-C₃ alkyl. In some embodiments, x is 1, y is 0 and R ⁷ is methyl.

In some embodiments, R ⁸ is Cl. In some embodiments, R ⁸ is —cn.

In some embodiments, R ⁹ is F.

In some embodiments, X ³ is N. In some embodiments, X ³ is CH.

In some embodiments, z is 0. In some embodiments, z is 1.

In some embodiments, R ^10a is H or halo. In some embodiments, R ^10a is H, F, cl, br or I.

In some embodiments, R ^10a is H.

In some embodiments, R ^10a is halo. In some embodiments, R ^10a is H, F, cl, br or I. In some embodiments, R ^10a is F.

In some embodiments, R ^10b is H or halo. In some embodiments, R ^10b is H, F, cl, br or I.

In some embodiments, R ^10b is H.

In some embodiments, R ^10b is halo. In some embodiments, R ^10b is H, F, cl, br or I. In some embodiments, R ^10b is F.

In some embodiments, R ¹¹ is H, C ₁-C₆ alkyl, - (C ₁-C₆ alkylene) - (5-to 6-membered heterocyclyl), - (C ₁-C₆ alkylene) -O (C ₁-C₆ alkyl), C ₁-C₆ haloalkyl, C ₁-C₆ alkyl-OH or- (C ₁-C₆ alkylene) -NH (C ₁-C₆ alkyl) wherein the heterocyclyl contains 1 to 3 groups selected from N, Heteroatoms of O and S. In some embodiments, R ¹¹ is H, C ₁-C₃ alkyl, - (C ₁-C₃ alkylene) - (5-to 6-membered heterocyclyl), - (C ₁-C₃ alkylene) -O (C ₁-C₃ alkyl), C ₁-C₅ haloalkyl, C ₁-C₅ alkyl-OH or- (C ₁-C₃ alkylene) -NH (C ₁-C₃ alkyl) wherein the heterocyclyl contains 1 to 3 groups selected from N, Heteroatoms of O and S. In some embodiments, R ¹¹ is H, C ₁-C₃ alkyl, - (C ₁-C₃ alkylene) - (5-to 6-membered heterocyclyl), - (C ₁-C₃ alkylene) -O (C ₁-C₃ alkyl), C ₁-C₅ haloalkyl, C ₁-C₅ alkyl-OH or- (C ₁-C₃ alkylene) -NH (C ₁-C₃ alkyl) wherein the heterocyclyl contains 1-2 heteroatoms selected from N and O. In some embodiments, R ¹¹ is H, methyl, ethyl, propyl, -CD ₃、-(CH₂) - (5-to 6-membered heterocyclyl), - (CH ₂CH₂) - (5-to 6-membered heterocyclyl), - (CH ₂CH₂CH₂) - (5-to 6-membered heterocyclyl )、-CH₂OCH₃、-CH₂OCH₂CH₃、-CH₂OCH₂CH₂CH₃、-CH₂CH₂OCH₃、-CH₂CH₂OCH₂CH₃、-CH₂CH₂OCH₂CH₂CH₃、-CH₂CH₂CH₂OCH₃、-CH₂CH₂CH₂OCH₂CH₃、-CH₂CH₂CH₂OCH₂CH₂CH₃、-(CH₂)₃CF₃、-(CH₂)₃CHF₂、-(CH₂)₃CH₂F、-CH₂CF₂CH₃、-CH₂CHFCH₃、-CH₂CF₂CH₃、-CH₂CH₂CF₃、-CH₂CH₂CF₂(CH₃)、-CH₂CH₂CF(CH₃)₂、-CH₂OH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CH₂CH(OH)(CH₃)、-CH₂CH₂C(OH)(CH₃)₂、-CH₂-N(H)CH₃、-CH₂-N(H)CH₂CH₃、-CH₂-N(H)CH₂CH₂CH₃、-CH₂CH₂-N(H)CH₃、-CH₂CH₂-N(H)CH₂CH₃、-CH₂CH₂-N(H)CH₂CH₂CH₃、-CH₂CH₂CH₂-N(H)CH₃、-CH₂CH₂CH₂-N(H)CH₂CH₃, or-CH ₂CH₂CH₂-N(H)CH₂CH₂CH₃, wherein the heterocyclyl contains 1-2 heteroatoms selected from N and O.

In some embodiments, R ¹¹ is H.

In some embodiments, R ¹¹ is C ₁-C₆ alkyl. In some embodiments, R ¹¹ is C ₁-C₃ alkyl. In some embodiments, R ¹¹ is methyl, ethyl, propyl, or-CD ₃. In some embodiments, R ¹¹ is-CH ₃. In some embodiments, R ¹¹ includes one or more deuterium atoms. In some embodiments, R ¹¹ is-CD ₃. In some embodiments, R ¹¹ is-CH ₂CH₃. In some embodiments, R ¹¹ is-CH (CH ₃)₂.

In some embodiments, R ¹¹ is- (C ₁-C₆ alkylene) - (5-to 6-membered heterocyclyl), wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S. In some embodiments, R ¹¹ is- (C ₁-C₃ alkylene) - (5-to 6-membered heterocyclyl), wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S. In some embodiments, R ¹¹ is- (C ₁-C₃ alkylene) - (5-to 6-membered heterocyclyl), wherein the heterocyclyl contains 1-2 heteroatoms selected from N and O. In some embodiments, R ¹¹ is。

In some embodiments, R ¹¹ is- (C ₁-C₆ alkylene) -O (C ₁-C₆ alkyl). In some embodiments, R ¹¹ is- (C ₁-C₃ alkylene) -O (C ₁-C₃ alkyl). In some embodiments, R ¹¹ is -CH₂OCH₃、-CH₂OCH₂CH₃、-CH₂OCH₂CH₂CH₃、-CH₂CH₂OCH₃、-CH₂CH₂OCH₂CH₃、-CH₂CH₂OCH₂CH₂CH₃、-CH₂CH₂CH₂OCH₃、-CH₂CH₂CH₂OCH₂CH₃ or-CH ₂CH₂CH₂OCH₂CH₂CH₃. In some embodiments, R ¹¹ is-CH ₂CH₂OCH₃.

In some embodiments, R ¹¹ is C ₁-C₆ haloalkyl. In some embodiments, R ¹¹ is C ₁-C₅ haloalkyl. In some embodiments, R ¹¹ is -CF₃、-CH₂CH₂F₃、-(CH₂)₂CHF₃、-(CH₂)₃CF₃、-CH₂CF(CH₃)₂、-CH₂CF₂CH₃、-CH₂CF₃、-CH₂CH₂CF(CH₃)₂、-CH₂CH₂CF₂CH₃ or-CH ₂CH₂CF₃. In some embodiments, R ¹¹ is -(CH₂)₃CF₃、-CH₂CF₂CH₃、-CH₂CH₂CF(CH₃)₂ or-CH ₂CF(CH₃)₂.

In some embodiments, R ¹¹ is C ₁-C₆ alkyl-OH. In some embodiments, R ¹¹ is C ₁-C₅ alkyl-OH. In some embodiments, R ¹¹ is -CH₂OH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CH₂CH(OH)(CH₃) or-CH ₂CH₂C(OH)(CH₃)₂. In some embodiments, R ¹¹ is-CH ₂CH₂C(OH)(CH₃)₂.

In some embodiments, R ¹¹ is- (C ₁-C₆ alkylene) -NH (C ₁-C₆ alkyl). In some embodiments, R ¹¹ is- (C ₁-C₃ alkylene) -NH (C ₁-C₃ alkyl). In some embodiments, R ¹¹ is -CH₂-N(H)CH₃、-CH₂-N(H)CH₂CH₃、-CH₂-N(H)CH₂CH₂CH₃、-CH₂CH₂-N(H)CH₃、-CH₂CH₂-N(H)CH₂CH₃、-CH₂CH₂-N(H)CH₂CH₂CH₃、-CH₂CH₂CH₂-N(H)CH₃、-CH₂CH₂CH₂-N(H)CH₂CH₃ or-CH ₂CH₂CH₂-N(H)CH₂CH₂CH₃. In some embodiments, R ¹¹ is-CH ₂CH₂N(H)CH₃.

。

In some embodiments, R ¹² is H, halo, or C ₁-C₆ alkyl. In some embodiments, R ¹² is H, F, cl, br, I or C ₁-C₃ alkyl. In some embodiments, R ¹² is H, F, cl, br, I, methyl, ethyl, or propyl.

In some embodiments, R ¹² is H.

In some embodiments, R ¹² is halo. In some embodiments, R ¹² is F, cl, br, or I. In some embodiments, R ¹² is F.

In some embodiments, R ¹² is C ₁-C₆ alkyl. In some embodiments, R ¹² is C ₁-C₃ alkyl. In some embodiments, R ¹² is methyl, ethyl, or propyl. In some embodiments, R ¹² is methyl.

。

in some embodiments, R ¹³ is H or halo. In some embodiments, R ¹³ is H, F, cl, br or I.

In some embodiments, R ¹³ is H.

In some embodiments, R ¹³ is halo. In some embodiments, R ¹³ is F, cl, br, or I. In some embodiments, R ¹³ is F.

In some embodiments, R ¹⁴ is H or C ₁-C₆ alkyl. In some embodiments, R ¹⁴ is H or C ₁-C₃ alkyl. In some embodiments, R ¹⁴ is H, methyl, ethyl, or propyl.

In some embodiments, R ¹⁴ is H.

In some embodiments, R ¹⁴ is C ₁-C₆ alkyl. In some embodiments, R ¹⁴ is C ₁-C₃ alkyl. In some embodiments, R ¹⁴ is methyl, ethyl, or propyl. In some embodiments, R ¹⁴ is methyl.

In some embodiments, R ¹⁴ is H. In some embodiments, R ¹⁴ is methyl.

In some embodiments, X ⁴ is N. In some embodiments, X ⁴ is CR ¹⁵.

In some embodiments, R ¹⁵ is H or C ₁-C₆ alkyl. In some embodiments, R ¹⁵ is H or C ₁-C₃ alkyl. In some embodiments, R ¹⁵ is H, methyl, ethyl, or propyl.

In some embodiments, R ¹⁵ is H.

In some embodiments, R ¹⁵ is C ₁-C₆ alkyl. In some embodiments, R ¹⁵ is C ₁-C₃ alkyl. In some embodiments, R ¹⁵ is methyl, ethyl, or propyl. In some embodiments, R ¹⁵ is methyl.

In some embodiments, X ⁵ is N. In some embodiments, X ⁵ is CH.

In some embodiments, X ⁶ is N. In some embodiments, X ⁶ is CH.

In some embodiments of the present invention, in some embodiments,Is a single bond. In some embodiments of the present invention, in some embodiments,Is a double bond.

。

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (II):

Wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁷、R⁸、R⁹、R^10a、R^10b、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IIa):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁷、R⁸、R⁹、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IIb):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁷、X¹、X² and w are as described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IIc):

Wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R⁷、X¹ and X ² are as described for formula (IA) or (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (III):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁶、R⁸、R⁹、R^10a、R^10b、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IIIa):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁶、R⁸、R⁹、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IIIb):

wherein R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b and R ¹¹ are as described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IV):

Wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁸、R⁹、R^10a、R^10b、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IVa):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁸、R⁹、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、X¹、X²、X³、X⁴、X⁵、X⁶、w、z and As described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IVb):

wherein R^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、X¹、X² and w are as described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IVc):

Wherein R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、X¹ and X ² are as described for formula (IA) or formula (I).

In some embodiments, the compound of formula (IA) or formula (I) is a compound of formula (IVd):

Wherein R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a and R ^4b are as described for formula (IA) or formula (I).

It is to be understood that any of the compounds described herein may include substitution of deuterium for one or more hydrogen atoms. Any one or more of the substituents of formula (IA) or formula (I) may be deuterated, such as one or more of X¹、X²、X³、X⁵、X⁶、R^1a、R^1b、R^3a、R^3b、R^4a、R^4b、R^5a、R^5b、R⁶、R⁷、R^10a、R^10b、R¹¹、R¹²、R¹³、R¹⁴ and R ¹⁵. For example, in some embodiments, R ¹¹ is a deuterated group, such as-CD ₃.

In the description herein, it is to be understood that all descriptions, variations, embodiments, or aspects of formula (IA) or formula (I) apply equally to other formulae detailed herein, where applicable, and are described equally as if each description, variation, embodiment, or aspect were separately and individually listed. It is also to be understood that all descriptions, variations, embodiments, or aspects of formula (IA) or formula (I) apply equally to other formulae detailed herein, where applicable, and are described equally as if each was separately and individually listed for all formulae. For example, all descriptions, variations, embodiments, or aspects of formula (IA) or formula (I) apply equally to any formula detailed herein, where applicable, such as formulas (II), (IIa), (IIb), (IIc), (III), (IIIa), (IIIb), (IV), (IVa), (IVb), (IVc), and (IVd), and are described equally as if each of the descriptions, variations, embodiments, or aspects were separately and individually listed for all formulas.

In some embodiments, provided are compounds selected from the compounds of table 1, or pharmaceutically acceptable salts thereof. Although certain compounds described in this disclosure (including table 1) are presented in particular stereoisomers and/or non-stereochemical forms, it should be understood that any or all stereochemical forms (including any enantiomeric or diastereomeric forms) and any tautomeric or other forms of any of the compounds in this disclosure (including table 1) are described herein.

Table 1.

Or a pharmaceutically acceptable salt thereof.

All compounds of formula (IA) or (I) present in free base or acid form can be converted into their pharmaceutically acceptable salts by methods known to the person skilled in the art by treatment with suitable inorganic or organic bases or acids. Salts of the compounds of formula (IA) or (I) may be converted to their free base or acid form by standard techniques.

Synthesis method

The compounds described herein can be prepared using conventional organic synthesis and commercially available starting materials or methods provided herein. By way of example and not limitation, compounds of formulas (I-a) through (I-h) may be prepared as outlined in schemes 1-4 and the examples set forth herein. It should be noted that those skilled in the art will know how to modify the procedures set forth in the exemplary schemes and examples to obtain the desired product.

As outlined in scheme 1, compounds of formula (i-1) containing an oxindole motif can be synthesized from indole derivative a coupled with intermediate compound b to form intermediate compound c, which is then nitrated to form intermediate compound d. The nitro group is reduced to form intermediate compound e, which is subsequently coupled with intermediate f to form the compound of formula (i-1).

Scheme 2 provides two routes for the synthesis of compounds of formula (i-2) containing glutarimide motifs. Indazole derivative g can be coupled with intermediate compound h to give intermediate compound i. Deprotection and further reduction of intermediate compound i forms intermediate compound j, which is subjected to deprotection to form a compound of formula (i-2). Alternatively, indazole derivative g can be coupled with intermediate compound k to give intermediate compound i. Deprotection and further reduction of intermediate compound l forms intermediate compound i, which is subjected to deprotection of an amine to form a compound of formula (i-2).

Scheme 3 provides for the synthesis of compounds of formula (i-2) containing a dihydrouridine motif. Indazole derivative n can be coupled with intermediate compound h to give intermediate compound o, which is deprotected to form intermediate compound p and subsequently deprotected to form a compound of formula (i-2).

Scheme 4 provides a variety of pathways for synthesizing various compounds of formula (IA) or formula (I). The compound of formula (I-1) may be coupled with various derivatives of the compound of formula (I-2) (represented by formula (I-2 a) to formula (I-2 h)) to give the compounds of formula (I-a) to formula (I-h).

Application method

Embodiments of the present disclosure provide a method for modulating BCL6 in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or (I). Modulation (e.g., inhibition or activation) of BCL6 can be assessed and demonstrated in a variety of ways known in the art. Kits and commercially available assays can be used to determine whether and to what extent BCL6 is modulated (e.g., inhibited or activated).

In one aspect, provided herein is a method of modulating BCL6, comprising contacting BCL6 with an effective amount of a compound of formula (IA) or formula (I), or any embodiment or variant thereof. In some embodiments, the compound of formula (IA) or formula (I) inhibits BCL6. In some embodiments, the compound of formula (IA) or formula (I) causes degradation of BCL6.

In some embodiments, a compound of formula (IA) or formula (I) modulates the activity of BCL6 by 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%. In some embodiments, the compound of formula (IA) or formula (I) modulates the activity of BCL6 by 1%-100%、5%-100%、10%-100%、15%-100%、20%-100%、25%-100%、30%-100%、35%-100%、40%-100%、45%-100%、50%-100%、55%-100%、60%-100%、65%-100%、70%-100%、75%-100%、80%-100%、85%-100%、90%-100%、95%-100%、5%-95%、5%-90%、5%-85%、5%-80%、5%-75%、5%-70%、5%-65%、5%-60%、5%-55%、5%-50%、5%-45%、5%-40%、5%-35%、5%-30%、5%-25%、5%-20%、5%-15%、5%-10%、10%-90%、20%-80%、30%-70% or 40% -60%.

Also provided in certain embodiments of the present disclosure is a method for degrading BCL6 in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or (I). Degradation of BCL6 can be assessed and demonstrated in a variety of ways known in the art. Kits and commercially available assays (including cell-based assays) can be used to determine whether and to what extent BCL6 has been degraded.

In one aspect, provided herein is a method of degrading BCL6, comprising contacting BCL6 with an effective amount of a compound of formula (IA) or formula (I), or any embodiment or variant thereof. In some embodiments, the compound of formula (IA) or formula (I) partially degrades BCL 6. In some embodiments, the compound of formula (IA) or formula (I) completely degrades BCL 6.

In some embodiments, a compound of formula (IA) or formula (I) degrades BCL6 by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the compound of formula (IA) or formula (I) degrades BCL6 by about 1%-100%、5%-100%、10%-100%、15%-100%、20%-100%、25%-100%、30%-100%、35%-100%、40%-100%、45%-100%、50%-100%、55%-100%、60%-100%、65%-100%、70%-100%、75%-100%、80%-100%、85%-100%、90%-100%、95%-100%、5%-95%、5%-90%、5%-85%、5%-80%、5%-75%、5%-70%、5%-65%、5%-60%、5%-55%、5%-50%、5%-45%、5%-40%、5%-35%、5%-30%、5%-25%、5%-20%、5%-15%、5%-10%、10%-90%、20%-80%、30%-70% or 40% -60%.

In another aspect, provided herein is a method for treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or (I). In some embodiments, provided herein is a method for preventing cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or (I). Non-limiting examples of cancers include squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma, renal cell carcinoma, bladder carcinoma, bowel cancer, breast cancer, cervical cancer, colon cancer, esophageal cancer, head cancer, kidney cancer, liver cancer, lung cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, leukemia, benign lymphoma, malignant lymphoma, burkitt's lymphoma, non-hodgkin lymphoma (NHL), benign melanoma, malignant melanoma, myeloproliferative disease, sarcoma, ewing's sarcoma, angiosarcoma, kaposi's sarcoma, liposarcoma, myosarcoma, peripheral nerve epithelial tumor, synovial sarcoma, glioma, astrocytoma, oligodendroglioma, ependymoma, glioblastoma, neuroblastoma, ganglion glioma medulloblastoma, pineal tumor, meningioma, neurofibroma and schwannoma, prostate cancer, uterine cancer, testicular cancer, thyroid cancer, astrocytoma, gastric cancer, melanoma, carcinoma sarcoma, hodgkin's disease, nephroblastoma, teratocarcinoma, T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, pre-B ALL, pre-B lymphoma, diffuse large B-cell lymphoma, B-cell ALL, philadelphia chromosome positive CML, follicular lymphoma, marginal zone lymphoma, mantle cell lymphoma, fahrenheit macroglobulinemia, chronic Lymphocytic Leukemia (CLL), small Lymphocytic Lymphoma (SLL), intravascular large B-cell lymphoma, perivascular large B-cell lymphoma, B-cell leukemia, chronic myeloid leukemia, and non-small cell lung cancer.

In some embodiments, the cancer is a lymphoma, such as a B cell lymphoma. In some embodiments, the cancer is non-hodgkin's lymphoma, such as B-cell non-hodgkin's lymphoma. In some embodiments, the lymphoma is diffuse large B-cell lymphoma (DLBCL), follicular Lymphoma (FL), mantle cell lymphoma, marginal zone lymphoma (e.g., mucosa-associated lymphoid tissue lymphoma, peri-nodal B-cell lymphoma, splenic marginal zone B-cell lymphoma), burkitt's lymphoma, or lymphoplasmacytic lymphoma (megaloblastic fahrenheit). In some embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL) or Follicular Lymphoma (FL). In some embodiments, the cancer is DLBCL (e.g., recurrent and/or refractory DLBCL). In some embodiments, the cancer is FL (e.g., recurrent and/or refractory FL).

In some embodiments, the cancer is recurrent and/or refractory.

In some embodiments, the compound of formula (IA) or (I) is administered as a first line therapy. In other embodiments, the compound of formula (IA) or (I) is administered as a non-first line therapy (e.g., as a second line or subsequent therapy, such as a rescue therapy).

In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof reduces the extent of cancer (such as tumor size, tumor growth rate, metastasis) in the subject. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof stabilizes (prevents or delays progression according to) the cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof delays the onset or recurrence of cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof slows progression of cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof provides partial remission of cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof provides complete remission of cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof reduces the dose of one or more other drugs required to treat cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof enhances the effect of another drug for treating cancer. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof delays progression of cancer. In some embodiments, administering a compound of formula (IA) or formula (I) to a subject in need thereof improves the quality of life of the subject suffering from cancer. In some embodiments, administering a compound of formula (IA) or formula (I) to a subject in need thereof increases the survival of a subject suffering from cancer.

In some aspects, provided herein is a method of slowing the progression of cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or (I). In some embodiments, provided herein is a method of stabilizing cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, the method prevents progression of cancer. In some embodiments, the method delays progression of cancer. In some embodiments, the methods provide partial or complete remission of cancer.

In another aspect, provided herein is a method of delaying the onset or recurrence of cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I).

In a further aspect, provided herein is a method of reducing the dose of one or more other drugs required to treat cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, provided herein is a method of enhancing the effect of another drug for treating cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I).

Also provided herein is a method of delaying progression of cancer in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, the methods improve the quality of life of a subject with cancer. In some embodiments, the methods extend the survival of a subject with cancer.

In a further aspect, provided herein is a method for treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or (I). In some embodiments, provided herein is a method for preventing an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA) or formula (I). Autoimmune diseases can be divided into two categories. Organ-specific autoimmune diseases occur when the immune system targets a particular cell, tissue or organ. A generalized autoimmune disease occurs when the immune system attacks the body without distinguishing between different types of tissues or target cells. Exemplary organ-specific autoimmune diseases include atopic dermatitis, asthma, insulin-dependent diabetes mellitus, hashimoto thyroiditis, graves 'disease, pernicious anemia, myasthenia gravis, pemphigus vulgaris, and crohn's disease. Exemplary generalized autoimmune diseases include Systemic Lupus Erythematosus (SLE), rheumatoid arthritis, scleroderma, sarcoidosis, and gillin-barre syndrome (GBS). The present disclosure encompasses the treatment of all types of autoimmune diseases, including organ-specific and pan-type autoimmune diseases, including, but not limited to, lupus erythematosus, ankylosing spondylitis, chagas's disease (CHAGAS DISEASE), chronic obstructive pulmonary disease, crohn's disease, dermatomyositis, type 1 diabetes, endometriosis, pulmonary hemorrhagic nephritis syndrome (Goodpasture's syndrome), graves's disease, gill-barre syndrome (GBS), hashimoto's disease, suppurative sweat gland, kawasaki disease, igA nephropathy, idiopathic thrombocytopenic purpura, interstitial cystitis, mixed connective tissue disease, scleroderma, multiple sclerosis, myasthenia gravis, narcolepsy, neuromuscular rigidity, pemphigus vulgaris, pernicious anemia, psoriasis, psoriatic arthritis, polymyositis, primary biliary cirrhosis, recurrent polychondritis, rheumatoid arthritis, sarcoidosis, psoriatic arthritis, polyarthritis, and vasculitis, and temporal disease.

In another aspect, provided herein is a method for treating a TH 17-related disorder, such as a TH 17-related autoimmune disorder, in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (IA) or (I).

In some embodiments, administration of a compound of formula (IA) or (I) to a subject in need thereof reduces or alleviates symptoms of an autoimmune disease in the subject (such as inflammation, chronic fever, weakness, joint pain, myalgia, and fatigue). In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof shortens or reduces the duration of symptoms of an autoimmune disease. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof eliminates symptoms of autoimmune disease. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof delays the onset or recurrence of an autoimmune disease. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof slows the progression of an autoimmune disease. In some embodiments, administration of a compound of formula (IA) or (I) to a subject in need thereof reduces the dosage of one or more other drugs required to treat an autoimmune disease. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof enhances the effect of another agent for treating an autoimmune disease. In some embodiments, administration of a compound of formula (IA) or formula (I) to a subject in need thereof delays the progression of an autoimmune disease. In some embodiments, administering a compound of formula (IA) or formula (I) to a subject in need thereof improves the quality of life of the subject suffering from an autoimmune disease.

In some aspects, provided herein is a method of slowing the progression of an autoimmune disease in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, provided herein is a method of stabilizing an autoimmune disease in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, the method prevents the progression of an autoimmune disease. In some embodiments, the method delays the progression of the autoimmune disease. In some embodiments, the method improves the quality of life of a subject suffering from an autoimmune disease.

In a further aspect, provided herein is a method of reducing the dose of one or more other drugs required to treat an autoimmune disease in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I). In some embodiments, provided herein is a method of enhancing the effect of another drug for treating an autoimmune disease in a subject, the method comprising administering to the subject a compound of formula (IA) or formula (I).

In some embodiments of any of the methods described herein, the compound of formula (IA) or (I) is administered during one or more treatment cycles (e.g., three or more treatment cycles, six or more treatment cycles, 12 or more treatment cycles, 15 or more treatment cycles, 18 or more treatment cycles, or 24 or more treatment cycles). In a particular embodiment, the treatment cycle is a 28 day treatment cycle.

In some embodiments, the compound of formula (IA) or (I) is administered daily on each day of a 28-day treatment cycle, e.g., once or twice daily on each day of a 28-day treatment cycle. In some embodiments, the compound of formula (IA) or (I) is administered daily on days 1-14 of the 28 day treatment cycle and is not administered on days 15-28 of the 28 day treatment cycle, e.g., the compound of formula (IA) or (I) is administered once or twice daily on days 1-14 of the 28 day treatment cycle. In some embodiments, the compound of formula (IA) or (I) is administered daily on days 1-21 of the 28 day treatment cycle and is not administered on days 22-28 of the 28 day treatment cycle, e.g., the compound of formula (IA) or (I) is administered once or twice daily on days 1-21 of the 28 day treatment cycle.

Administration of the compound of formula (IA) or (I) may continue indefinitely according to the methods described herein, or until a clinically significant disease progression, unacceptable toxicity, or decision to otherwise cease administration is reached. In some embodiments, administration of the compound of formula (IA) or (I) continues for up to three years, e.g., up to two years, and then ceases.

In some embodiments of any of the methods described herein, a compound of formula (IA) or (I) is administered in combination with an additional therapy. Thus, some embodiments further comprise administering to the subject an additional therapy, such as an additional therapeutic agent. In some embodiments, the additional therapy is standard of care (SOC).

Current SOCs for DLBCL are the combined administration of CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) with rituximab (R-CHOP). SOC for secondary recurrent/refractory DLBCL is high dose chemotherapy followed by autologous stem cell transplantation (autoSCT). CAR-T cell therapies have also been used to treat recurrent/refractory DLBCL. SOC for FL is anti-CD 20 immunotherapy with or without chemotherapy.

In some embodiments, the additional therapy comprises an anti-lymphoma therapy (e.g., an anti-lymphoma agent). Anti-lymphoma therapies include the SOC described herein for DLBCL and FL, as well as cytotoxic agents, radiation therapy, radioimmunotherapy, and autologous and allogeneic grafts. Anti-lymphoma agents include SOC agents described herein for DLBCL and FL (e.g., cyclophosphamide, doxorubicin, vincristine, prednisolone, rituximab), as well as cytotoxic agents and anti-CD 20 antibodies.

Rituximab is a recombinant chimeric murine/human antibody directed against the CD20 antigen, a hydrophobic transmembrane protein located on normal pre-B lymphocytes and mature B lymphocytes. Following binding, rituximab triggered a cytotoxic immune response against CD20 positive cells. Rituximab is indicated for a variety of tumor treatments and non-tumor treatments. For example, rituximab is indicated for (i) treating patients with previously untreated diffuse large B-cell CD20 positive NHL in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) or other anthracycline-based chemotherapy regimens, (ii) treating patients with previously untreated follicular CD20 positive B-cell NHL in combination with CVP (cyclophosphamide, vincristine, prednisolone) chemotherapy, and/or (iii) treating patients with recurrent or refractory, low-grade or follicular, CD20 positive, B-cell NHL as a single agent. For example, rituximab is indicated for (i) treating patients with CD20 positive diffuse large B-cell non-Hodgkin's lymphoma in combination with CHOP chemotherapy, (ii) treating patients with previously untreated stage III-IV FL in combination with chemotherapy, (III) treating patients with stage III-IV FL that are resistant to chemotherapy or are in their second or subsequent relapse after chemotherapy as monotherapy, and/or (IV) treating adult patients with FL that are responsive to induction therapy as maintenance therapy. Rituximab is also a component of most salvage chemotherapy regimens (e.g., R-ICE, R-DHAP, R-esap, R-gemcitabine/oxaliplatin) and is listed in NCCN and european medical institute (European Society of Medical Oncology, ESMO) guidelines as a treatment option for two-wire (and subsequent) treatment of B-cell lymphomas, and is routinely used in standard practice in these contexts. Furthermore, rituximab is indicated for use in combination with lenalidomide for the treatment of relapsed/refractory FL in the united states and the european union.

In some embodiments, the method further comprises administering an anti-CD 20 antibody (e.g., a therapeutically effective amount of an anti-CD 20 antibody), such as rituximab, to the subject. In particular embodiments, the method further comprises administering rituximab (e.g., a therapeutically effective amount of rituximab) to the subject. For example, some embodiments include intravenous administration of rituximab (e.g., as an Intravenous (IV) infusion). Some embodiments include administering about 375 mg/m ² rituximab. Some embodiments include intravenous (e.g., as an IV infusion) administration of about 375 mg/m ² rituximab. In some embodiments, rituximab is administered simultaneously with or within up to 2 hours after the compound of formula (IA) or (I). Some embodiments include administering rituximab once every 28 days (e.g., on day 1 of a 28 day cycle). In some embodiments, rituximab is administered once every 28 days (e.g., on day 1 of a 28 day cycle) for up to 12 months, e.g., for up to 6 months.

Pharmaceutical compositions and routes of administration

The compounds provided herein can be administered to a subject orally, topically, or parenterally in the form of conventional formulations, such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions.

The compounds disclosed herein may be administered to a subject orally, topically or parenterally in the form of conventional formulations, such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions and emulsions. Suitable formulations may be prepared by methods commonly employed using conventional organic or inorganic additives such as excipients (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate or calcium carbonate), binders (e.g., cellulose, methylcellulose, hydroxymethyl cellulose, polypropylene-pyrrolidone, polyvinylpyrrolidone, gelatin, acacia, polyethylene glycol, sucrose or starch), disintegrants (e.g., starch, carboxymethyl cellulose, hydroxypropyl starch, low substituted hydroxypropyl cellulose, sodium bicarbonate, calcium phosphate or calcium citrate), lubricants (e.g., magnesium stearate, light anhydrous silicic acid, talc or sodium lauryl sulfate), flavoring agents (e.g., citric acid, menthol, glycine or orange powder), preservatives (e.g., sodium benzoate, sodium bisulphite, methyl or propyl parahydroxybenzoate), stabilizers (e.g., citric acid, sodium citrate or acetic acid), suspending agents (e.g., methylcellulose, polyvinylpyrrolidone or aluminum stearate), dispersants (e.g., hydroxypropyl methylcellulose), diluents (e.g., polyethylene glycol, and waxes, such as may be petrolatum, or white petrolatum. An effective amount of a compound of formula (IA) or formula (I) in a pharmaceutical composition may be at a level that will exert the desired effect, for example, from about 0.005 mg/kg subject body weight to about 10 mg/kg subject body weight in unit dose for both oral and parenteral administration.

The dosage of the compound of formula (IA) or (I) to be administered to a subject is quite widely variable and may depend on the judgment of the health care practitioner. Generally, the compounds disclosed herein can be administered one to four times per day at a dose of about 0.001 mg/kg of subject body weight to about 10 mg/kg of subject body weight, although the above dose can vary as appropriate depending on the age, weight, and medical condition of the subject and the type of administration. In one embodiment, the dose is from about 0.001 mg/kg subject weight to about 5 mg/kg subject weight, from about 0.01 mg/kg subject weight to about 5 mg/kg subject weight, from about 0.05 mg/kg subject weight to about 1 mg/kg subject weight, from about 0.1 mg/kg subject weight to about 0.75 mg/kg subject weight, or from about 0.25 mg/kg subject weight to about 0.5 mg/kg subject weight. In one embodiment, one dose is administered daily. In any given case, the amount of compound of formula (IA) or (I) administered will depend on factors such as the solubility of the active ingredient, the formulation used and the route of administration.

In some embodiments, the compound of formula (IA) or formula (I) is administered to the subject at a dose of about 0.01 mg/day to about 750 mg/day, about 0.1 mg/day to about 375 mg/day, about 0.1 mg/day to about 150 mg/day, about 0.1 mg/day to about 75 mg/day, about 0.1 mg/day to about 50 mg/day, about 0.1 mg/day to about 25 mg/day, or about 0.1 mg/day to about 10 mg/day. In some embodiments, the compound of formula (IA) or formula (I) is administered to the subject once daily or twice daily at a dose of about 10 mg/day to about 750 mg/day.

In particular embodiments, the compound of formula (IA) or (I) is administered to the subject at a dose of about 20 mg/day, about 40 mg/day, about 80 mg/day, about 160 mg/day, about 260 mg/day, about 400 mg/day, or about 640 mg/day once daily (QD) or twice daily (BID). In further embodiments, the administration is QD. Alternatively, the administration is BID.

In another embodiment, provided herein are unit dose formulations comprising a compound of formula (IA) or formula (I) between about 0.1 mg and 500 mg, about 1 mg and 250 mg, about 1 mg and about 100mg, about 1 mg and about 50 mg, about 1 mg and about 25mg, or between about 1 mg and about 10 mg.

In a particular embodiment, provided herein are unit dose formulations comprising about 0.1 mg or 100mg of a compound of formula (IA) or formula (I). In some embodiments, the unit dose formulation comprises from about 10 mg to about 100mg of a compound of formula (IA) or formula (I). In some embodiments, the unit dose formulation comprises about 20 mg of a compound of formula (IA) or (I) (e.g., compound 70 or a pharmaceutically acceptable salt thereof). In some embodiments, the unit dose formulation comprises about 80 mg of a compound of formula (IA) or (I) (e.g., compound 70 or a pharmaceutically acceptable salt thereof).

In another embodiment, provided herein is a unit dose formulation comprising 0.5 mg、1 mg、5 mg、10 mg、15 mg、20 mg、30 mg、35 mg、50 mg、70 mg、100 mg、125 mg、140 mg、175 mg、200 mg、250 mg、280 mg、350 mg、500 mg、560 mg、700 mg、750 mg、1000 mg or 1400 mg of a compound of formula (IA) or formula (I).

The compound of formula (IA) or (I) may be administered once, twice, three times, four times or more daily. In particular embodiments, a dose of 100 mg or less is administered as a once daily dose, and a dose greater than 100 mg is administered twice daily in an amount equal to half the total daily dose. In some embodiments, the compound of formula (IA) or (I) is administered once daily. In some embodiments, the compound of formula (IA) or (I) is administered twice daily.

The compounds of formula (IA) or (I) may be administered orally. In one embodiment, the compound of formula (IA) or formula (I) is administered with a meal and water when administered orally. In another embodiment, the compound of formula (IA) or formula (I) is dispersed in water or fruit juice (e.g., apple or orange juice) or any other liquid and administered orally as a solution or suspension. Alternatively, in one embodiment, the compound of formula (IA) or (I) is administered to a subject in a fasted state when administered orally. For example, in some embodiments, a compound of formula (IA) or (I) is administered to a subject that does not eat or drink anything other than water at least about 2 hours prior to administration of the compound of formula (IA) or (I). In some embodiments, the fasted state begins at least about 2 hours prior to administration of the compound of formula (IA) or (I) and continues for at least about 1 hour after administration of the compound of formula (IA) or (I).

The compounds disclosed herein may also be administered intradermally, intramuscularly, intraperitoneally, transdermally, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, transdermally, rectally, transmucosally, by inhalation, or topically to the ear, nose, eye, or skin. The manner of administration is determined by the health care practitioner and may depend in part on the site in the medical condition.

In another embodiment, provided herein is a composition comprising an effective amount of a compound of formula (IA) or (I) and a pharmaceutically acceptable carrier or vehicle, wherein the pharmaceutically acceptable carrier or vehicle may comprise an excipient, diluent, or mixture thereof. In one embodiment, the composition is a pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprises a compound of formula (IA) or (I) and two or more of anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, and sodium stearyl fumarate. In some embodiments, the pharmaceutical composition comprises anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, and sodium stearyl fumarate.

In some embodiments, the composition is in the form of a dispersion (e.g., a spray-dried dispersion). The composition may optionally be coated with a film that resists dissolution, for example, for a predictable period of time.

The compound of formula (IA) or (I) may be present in the composition in any effective amount. In some embodiments, the compound of formula (IA) or (I) is present in the composition at 1% weight/weight (w/w) to 75% w/w, 5% w/w to 50% w/w, 10% w/w to 40% w/w, 30% w/w to 40% w/w, about 5% w/w, about 10% w/w, about 15% w/w, about 20% w/w, about 25% w/w, about 30% w/w, about 35% w/w, about 40% w/w, about 45% w/w, or about 50% w/w. In a particular embodiment, the pharmaceutical composition comprises about 35% w/w of the compound of formula (IA) or (I).

The compositions may be in the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, troches, suppositories, and suspensions. In some embodiments, the composition is in the form of a tablet or capsule. In some embodiments, the composition is in the form of a tablet (e.g., a film coated tablet). In some embodiments, the composition is in the form of a capsule.

The composition may be formulated to contain a daily dose or a suitable fraction of a daily dose in a dosage unit, which may be a single tablet or capsule or a suitable volume of liquid. In one embodiment, the solution is prepared from a water soluble salt such as the hydrochloride salt. In general, all compositions are prepared according to methods known in pharmaceutical chemistry. Capsules may be prepared by mixing a compound of formula (IA) or (I) with a suitable carrier or diluent and filling the appropriate amount of the mixture into capsules. Common carriers and diluents include, but are not limited to, inert powdered substances such as many different kinds of starches, powdered celluloses, especially crystalline celluloses and microcrystalline celluloses, sugars such as fructose, mannitol and sucrose, cereal flours and similar edible powders.

Tablets may be prepared by direct compression, wet granulation, dry granulation or spray drying. Their formulations typically incorporate diluents, binders, lubricants and disintegrants and compounds. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride, and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders are such substances as starch, gelatin and sugars such as lactose, fructose, glucose and the like. Natural gums and synthetic gums are also suitable, including acacia, alginate, methylcellulose, polyvinylpyrrolidone and the like. Polyethylene glycol, ethylcellulose, and waxes may also be used as binders.

Lubricants may be required in the tablet formulation to prevent sticking of the tablet and punch to the dye. The lubricant may be selected from the group consisting of slippery solids such as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils. Tablet disintegrants are substances that swell when wetted to break up the tablet and release the compound. They include starches, clays, celluloses, algins and gums. More specifically, for example, corn starch and potato starch, methyl cellulose, agar, bentonite, lignocellulose, powdered natural sponge, cation exchange resins, alginic acid, guar gum, citrus pulp and carboxymethyl cellulose and sodium lauryl sulfate may be used. Tablets may be coated with sugar as a flavoring and sealing agent, or with a film-forming protective agent to alter the dissolution characteristics of the tablet. The composition may also be formulated as a chewable tablet, for example by using a substance such as mannitol in the formulation.

When it is desired to administer a compound of formula (IA) or (I) as a suppository, a typical matrix may be used. Cocoa butter is a traditional suppository base that may be modified by the addition of waxes to slightly raise its melting point. Water-miscible suppository bases comprising polyethylene glycols of various molecular weights in particular are widely used.

The effect of the compounds of formula (IA) or (I) may be delayed or prolonged by appropriate formulation. For example, slow-dissolving pellets of a compound of formula (IA) or (I) may be prepared and incorporated into tablets or capsules, or incorporated as slow-release implantable devices. The technique also includes preparing pellets having several different dissolution rates and filling the capsule with a pellet mixture. Tablets or capsules may be coated with a film that resists dissolution over a predictable period of time. Parenteral formulations may even be made long-acting by dissolving or suspending a compound of formula (IA) or formula (I) in an oily or emulsifying vehicle which slowly disperses the compound in the serum. In some embodiments, a tablet or capsule is formulated for enteric release, for example, by formulating a compound of formula (IA) or (I) with Eudragit L100-55.

It is to be understood that the pharmaceutical compositions described herein may comprise mixtures of compounds of formula (IA) or (I), including racemic mixtures of any of the compounds described herein.

Numbering plan

Compounds useful in accordance with the present disclosure are further described by the following embodiments. The features of each embodiment may be combined with any other embodiment where appropriate and practical.

Embodiment p 1.A compound of formula (IA):

(IA)

or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

Embodiment p2. A compound of formula (I):

(I)

or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

Embodiment P3 the compound according to embodiment P1 or P2 or a pharmaceutically acceptable salt thereof, wherein:

x ¹ is CH, and

X ² is N.

Embodiment P4. The compound according to embodiment P1 or P2, or a pharmaceutically acceptable salt thereof, wherein:

x ¹ is N, and

X ² is CH.

Embodiment P5. the compound of embodiment P1 or P2, or a pharmaceutically acceptable salt thereof, wherein:

X ¹ and X ² are each N.

Embodiment P6. The compound according to any one of embodiments P1-P5, or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₃ alkyl.

Embodiment P7 the compound of embodiment P6 or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H or-CH ₃.

Embodiment P8 the compound according to any one of embodiments P1-P7, or a pharmaceutically acceptable salt thereof, wherein:

R ^2a and R ^2b are each independently H, C ₁-C₃ alkyl, -O (C ₁-C₃ alkyl), -OH, halo, C ₁-C₃ haloalkyl or C ₁-C₃ alkyl-OH,

Or R ^2a and R ^2b together with the carbon atom to which they are attached form a spiro C ₃-C₄ cycloalkyl,

Or R ^2a and R ^2b together form an oxo group.

Embodiment P9 the compound of embodiment P8 or a pharmaceutically acceptable salt thereof, wherein:

R ^2a and R ^2b are each independently H, -CH ₃、-OCH₃、-OH、F、-CF₃ or-CH ₂ OH,

Or R ^2a and R ^2b together with the carbon atom to which they are attached form a spirocyclobutyl group,

Or R ^2a and R ^2b together form an oxo group.

Embodiment P10 the compound according to any one of embodiments P1-P9, or a pharmaceutically acceptable salt thereof, wherein R ^1a and R ^1b are each H, and at least one of R ^2a and R ^2b is not H.

Embodiment P11 the compound according to any one of embodiments P1-P9 or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^2a together form a bridging ethylene group.

Embodiment P12 the compound according to any one of embodiments P1-P11, or a pharmaceutically acceptable salt thereof, wherein:

R ^3a and R ^3b are each independently H, halo or C ₁-C₃ alkyl.

Embodiment P13 the compound of embodiment P12 or a pharmaceutically acceptable salt thereof, wherein:

R ^3a and R ^3b are each independently H, F or-CH ₃.

Embodiment P14 the compound according to any one of embodiments P1-P13, or a pharmaceutically acceptable salt thereof, wherein:

R ^4a and R ^4b are each independently H, halo or C ₁-C₃ alkyl,

Or R ^4a and R ^4b together with the carbon atom to which they are attached form a spiro C ₃-C₄ cycloalkyl group.

Embodiment P15 the compound of embodiment P14 or a pharmaceutically acceptable salt thereof, wherein:

R ^4a and R ^4b are each independently H, F or-CH ₃,

Or R ^4a and R ^4b together with the carbon atom to which they are attached form a spirocyclobutyl group.

Embodiment P16 the compound of any one of embodiments P1-P15, or a pharmaceutically acceptable salt thereof, wherein R ^3a、R^3b、R^4a and R ^4b are each H.

Embodiment P17 the compound of any one of embodiments P1-P15, or a pharmaceutically acceptable salt thereof, wherein at least one of R ^3a and R ^3b is not H and at least one of R ^4a and R ^4b is not H.

Embodiment P18 the compound according to any one of embodiments P1-P17, or a pharmaceutically acceptable salt thereof, wherein:

w is 0.

Embodiment P19 the compound according to any one of embodiments P1-P17 or a pharmaceutically acceptable salt thereof, wherein:

w is 1.

Embodiment P20 the compound of embodiment P19 or a pharmaceutically acceptable salt thereof, wherein:

R ^5a and R ^5b are each independently H, halo or C ₁-C₃ alkyl.

Embodiment P21 the compound of embodiment P20 or a pharmaceutically acceptable salt thereof, wherein:

R ^5a and R ^5b are each H.

Embodiment P22 the compound according to any one of embodiments P1-P21, or a pharmaceutically acceptable salt thereof, wherein at least one of R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a and R ^5b is not H.

Embodiment P23 the compound of embodiment P22, or a pharmaceutically acceptable salt thereof, wherein one or both of R ^1a、R^1b、R^2a、R^2b、R^3a、R^3b、R^4a、R^4b、R^5a and R ^5b is not H.

Embodiment P24 the compound according to any one of embodiments P1 to P23 or a pharmaceutically acceptable salt thereof, whereinThe method comprises the following steps:

。

embodiment P25 the compound according to any one of embodiments P1-P24, or a pharmaceutically acceptable salt thereof, wherein:

x is 1;

y is 0, and

R ⁷ is H or C ₁-C₃ alkyl.

Embodiment P26 the compound of embodiment P25 or a pharmaceutically acceptable salt thereof, wherein:

R ⁷ is H or-CH ₃.

Embodiment P27 the compound according to any one of embodiments P1-P24 or a pharmaceutically acceptable salt thereof, wherein:

x is 0;

y is 1, and

R ⁶ is H or C ₁-C₃ alkyl.

Embodiment P28 the compound of embodiment P27 or a pharmaceutically acceptable salt thereof, wherein:

R ⁶ is H or-CH ₃.

Embodiment P29 the compound according to any one of embodiments P1-P24, or a pharmaceutically acceptable salt thereof, wherein:

x and y are each 0.

Embodiment P30 the compound according to any one of embodiments P1-P29, or a pharmaceutically acceptable salt thereof, wherein:

R ⁸ is Cl.

Embodiment P31 the compound according to any one of embodiments P1-P29, or a pharmaceutically acceptable salt thereof, wherein:

r ⁸ is-CN.

Embodiment P32 the compound according to any one of embodiments P1-P31, or a pharmaceutically acceptable salt thereof, wherein:

X ³ is CH.

Embodiment P33 the compound according to any one of embodiments P1-P31 or a pharmaceutically acceptable salt thereof, wherein:

X ³ is N.

Embodiment P34 the compound according to any one of embodiments P1-P33, or a pharmaceutically acceptable salt thereof, wherein:

z is 0.

Embodiment P35 the compound according to any one of embodiments P1-P33, or a pharmaceutically acceptable salt thereof, wherein:

z is 1.

Embodiment P36 the compound according to any one of embodiments P1-P35, or a pharmaceutically acceptable salt thereof, wherein:

R ^10a and R ^10b are each independently H or F.

Embodiment P37 the compound of any one of embodiments P1-P36, or a pharmaceutically acceptable salt thereof, wherein:

R ¹¹ is H, C ₁-C₃ alkyl, - (C ₁-C₃ alkylene) - (6 membered heterocyclyl), - (C ₁-C₃ alkylene) -O (C ₁-C₃ alkyl), C ₁-C₅ haloalkyl, C ₁-C₅ alkyl-OH or- (C ₁-C₃ alkylene) -NH (C ₁-C₃ alkyl),

Wherein the heterocyclic group contains 1 to 2 heteroatoms selected from N and O.

Embodiment P38 the compound of embodiment P37 or a pharmaceutically acceptable salt thereof, wherein:

R ¹¹ is H、-CH₃、-CD₃、-CH₂CH₃、-CH(CH₃)₂、-CH₂CH₂OCH₃、-CH₂CH₂OH、-CH₂CH₂C(OH)(CH₃)₂、-CH₂CH₂N(H)CH₃、-(CH₂)₃CF₃、-CH₂CF₂CH₃、-CH₂CH₂CF(CH₃)₂、-CH₂CF(CH₃)₂ or 。

Embodiment P39 the compound according to any one of embodiments P1-P38, or a pharmaceutically acceptable salt thereof, whereinThe method comprises the following steps:

。

Embodiment P40 the compound according to any one of embodiments P1-P39, or a pharmaceutically acceptable salt thereof, wherein:

r ¹² is H, halo or C ₁-C₃ alkyl.

Embodiment P41 the compound of embodiment P40 or a pharmaceutically acceptable salt thereof, wherein:

R ¹² is H, F or-CH ₃.

Embodiment P42 the compound according to any one of embodiments P1 to P41 or a pharmaceutically acceptable salt thereof, whereinThe method comprises the following steps:

。

Embodiment P43 the compound according to any one of embodiments P1-P42 or a pharmaceutically acceptable salt thereof, wherein:

r ¹³ is H or F.

Embodiment P44 the compound according to any one of embodiments P1-P43 or a pharmaceutically acceptable salt thereof, wherein:

R ¹⁴ is H or C ₁-C₃ alkyl.

Embodiment P45 the compound of embodiment P44 or a pharmaceutically acceptable salt thereof, wherein:

R ¹⁴ is H or-CH ₃.

Embodiment P46 the compound of any one of embodiments P1-P45 or a pharmaceutically acceptable salt thereof, wherein:

X ⁴ is N.

Embodiment P47 the compound according to any one of embodiments P1-P45 or a pharmaceutically acceptable salt thereof, wherein:

X ⁴ is CR ¹⁵, and

R ¹⁵ is H or C ₁-C₃ alkyl.

Embodiment P48 the compound of embodiment P47 or a pharmaceutically acceptable salt thereof, wherein:

X ⁴ is CR ¹⁵, and

R ¹⁵ is H or-CH ₃.

Embodiment P49 the compound according to any one of embodiments P1-P48 or a pharmaceutically acceptable salt thereof, wherein:

X ⁵ is N.

Embodiment P50 the compound according to any one of embodiments P1-P48 or a pharmaceutically acceptable salt thereof, wherein:

X ⁵ is CH.

Embodiment P51 the compound according to any one of embodiments P1-P50 or a pharmaceutically acceptable salt thereof, wherein:

X ⁶ is N.

Embodiment P52 the compound according to any one of embodiments P1-P50 or a pharmaceutically acceptable salt thereof, wherein:

x ⁶ is CH.

Embodiment P53 the compound according to any one of embodiments P1-P52 or a pharmaceutically acceptable salt thereof, wherein:

Is a single bond.

Embodiment P54 the compound according to any one of embodiments P1-P52, or a pharmaceutically acceptable salt thereof, wherein:

Is a double bond.

Embodiment P55 the compound according to any one of embodiments P1 to P54, or a pharmaceutically acceptable salt thereof, whereinThe method comprises the following steps:

。

Embodiment P56 the compound of any one of embodiments P1-P55, or a pharmaceutically acceptable salt thereof, wherein the compound has formula (II), (III), or (IV):

。

embodiment P57 a compound according to embodiment P56, or a pharmaceutically acceptable salt thereof, wherein the compound has formula (IIIb):

(IIIb)。

embodiment p58 a compound selected from the group consisting of the compounds of table 1 and pharmaceutically acceptable salts thereof.

Embodiment P59 a pharmaceutical composition comprising a compound according to any one of embodiments P1-P58, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

Embodiment P60. A method of degrading B cell lymphoma 6 protein (BCL 6), said method comprising contacting BCL6 with an effective amount of a compound according to any one of embodiments P1-P58 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to embodiment P59.

Embodiment P61 a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound according to any one of embodiments P1-P58 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to embodiment P59.

Embodiment P62 the method of embodiment P61, wherein the cancer is lymphoma.

Embodiment P63 the method of embodiment P62, wherein the lymphoma is diffuse large B-cell lymphoma.

Embodiment P64 a method of treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound according to any one of embodiments P1-P58 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to embodiment P59.

Embodiment P65 the method according to embodiment P64, wherein the autoimmune disease is selected from the group consisting of atopic dermatitis, asthma, lupus erythematosus, ankylosing spondylitis, chagas's disease, chronic obstructive pulmonary disease, crohn's disease, dermatomyositis, type 1 diabetes, endometriosis, pulmonary hemorrhagic nephritis syndrome, graves 'disease, gillin-barre syndrome (GBS), hashimoto's disease, hidradenitis suppurativa, kawasaki disease, igA nephropathy, idiopathic thrombocytopenic purpura, interstitial cystitis, mixed connective tissue disease, scleroderma, multiple sclerosis, myasthenia gravis, narcolepsy, neuromuscular rigidity, pemphigus vulgaris, pernicious anemia, psoriasis, psoriatic arthritis, polymyositis, primary biliary cirrhosis, recurrent polychondritis, rheumatoid arthritis, sarcoidosis, schizophrenia, scleroderma, sjogren's syndrome, vitiligo, stiff person syndrome, vasculitis, granulomatosis, and wegiar's disease.

Embodiment p66 a pharmaceutical composition comprising a compound of the formula:

(70),

Or an isomer thereof, or a pharmaceutically acceptable salt of the foregoing, and two or more of anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, and sodium stearyl fumarate.

Embodiment P67 the pharmaceutical composition of embodiment P66, wherein the compound is

,

Or a pharmaceutically acceptable salt thereof.

Embodiment P68 the pharmaceutical composition of embodiment P66 or P67, comprising anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, and sodium stearyl fumarate.

Embodiment P69 the pharmaceutical composition according to any one of embodiments P66-P68, comprising about 35% weight/weight (w/w) of compound (70) or an isomer thereof, or a pharmaceutically acceptable salt of the foregoing.

Embodiment P70 the pharmaceutical composition according to any one of embodiments P66-P69, formulated for oral administration.

Embodiment P71 the pharmaceutical composition of embodiment P70, formulated for enteric release.

Embodiment P72. A unit dose formulation comprising the pharmaceutical composition according to any one of embodiments P66-P71, wherein the compound (70) or an isomer thereof or a pharmaceutically acceptable salt of the foregoing is present in the unit dosage form in an amount of from about 10mg to about 100 mg.

Embodiment P73 the unit dose formulation of embodiment P72 comprising about 20 mg or about 80 of the compound (70) or an isomer thereof of mg, or a pharmaceutically acceptable salt of the foregoing.

Embodiment P74 the unit dose formulation of embodiment P72 or P73, which is in the form of a tablet.

Embodiment P75 the unit dose formulation of embodiment P74, wherein the tablet is film coated.

Embodiment P76 a method of treating cancer or an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of an anti-CD 20 antibody and a compound according to any one of embodiments P1-P58, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of embodiments P59 and P66-P71, or a unit dose formulation according to any one of embodiments P72-P75.

Embodiment P77. A method of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of embodiments P1-P58, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of embodiments P59 and P66-P71, or a unit dose formulation according to any one of embodiments P72-P75,

Wherein the compound or pharmaceutically acceptable salt thereof is administered (i) daily on each day of a 28-day cycle, or (ii) on days 1-14 of a 28-day cycle and not on days 15-28 of the 28-day cycle, or (iii) on days 1-21 of a 28-day cycle and not on days 22-28 of the 28-day cycle.

Embodiment P78. A method of treating cancer or an autoimmune disease in a subject in need thereof, the method comprising administering to the subject the compound of any one of embodiments P1-P58, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of embodiments P59 and P66-P71, or the unit dose formulation of any one of embodiments P72-P75,

Wherein the compound or pharmaceutically acceptable salt thereof is administered to the subject at a dose of about 20 mg/day, about 40 mg/day, about 80 mg/day, about 160 mg/day, about 260 mg/day, about 400 mg/day, or about 640 mg/day once daily or twice daily.

Embodiment P79 the method according to embodiment P76 or P77, wherein the compound or pharmaceutically acceptable salt thereof is administered to the subject once daily or twice daily at a dose of about 20 mg/day, about 40 mg/day, about 80 mg/day, about 160 mg/day, about 260 mg/day, about 400 mg/day, or about 640 mg/day.

Embodiment P80 the method of any one of embodiments P76-P79, wherein the compound or pharmaceutically acceptable salt thereof is administered to the subject once daily.

Embodiment P81 the method of any one of embodiments P76-P79, wherein the compound or pharmaceutically acceptable salt thereof is administered to the subject twice daily.

Embodiment P82 the method according to any one of embodiments P76-P81, wherein the compound or pharmaceutically acceptable salt thereof is administered (i) daily on each day of a 28-day cycle, or (ii) on days 1-14 of a 28-day cycle and not on days 15-28 of the 28-day cycle, or (iii) on days 1-21 of a 28-day cycle and not on days 22-28 of the 28-day cycle.

Embodiment P83 the method of embodiment P82, wherein the compound or pharmaceutically acceptable salt thereof is administered daily on each day of a 28 day cycle.

Embodiment P84 the method of embodiment P82, wherein the compound or pharmaceutically acceptable salt thereof is administered on days 1-14 of a 28 day cycle and not administered on days 15-28 of the 28 day cycle.

Embodiment P85 the method of embodiment P82, wherein the compound or pharmaceutically acceptable salt thereof is administered on days 1-21 of a 28 day cycle and the compound is not administered on days 22-28 of the 28 day cycle.

Embodiment P86 the method of any one of embodiments P77-P85, further comprising administering to the subject an anti-CD 20 antibody.

Embodiment P87 the method of embodiment P76 or P86, wherein the anti-CD 20 antibody is rituximab.

Embodiment P88 the method of embodiment P87, wherein rituximab is administered intravenously.

Embodiment P89 the method of embodiment P87 or P88, wherein about 375 mg/m ² rituximab is administered once every 28 days.

Embodiment P90 the method according to any one of embodiments P87-P89, wherein rituximab is administered on day 1 of a 28-day treatment cycle and the compound or pharmaceutically acceptable salt thereof is administered (i) daily on each day of the 28-day treatment cycle, or (ii) on days 1-14 of the 28-day treatment cycle and not on days 15-28 of the 28-day treatment cycle, or (iii) on days 1-21 of the 28-day treatment cycle and not on days 22-28 of the 28-day treatment cycle.

Embodiment P91 the method of any one of embodiments P76-P90, wherein the method is a method of treating cancer.

Embodiment P92 the method of embodiment P91, wherein the cancer is lymphoma.

Embodiment P93 the method of embodiment P92, wherein the lymphoma is B-cell lymphoma.

Embodiment P94 the method according to embodiment P92 or P93, wherein the lymphoma is non-hodgkin's lymphoma.

Embodiment P95 the method according to any one of embodiments P92-P94, wherein the lymphoma is diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, burkitt's lymphoma, or lymphoplasmacytic lymphoma.

Embodiment P96 the method of embodiment P95, wherein the lymphoma is diffuse large B-cell lymphoma or follicular lymphoma.

Embodiment P97 the method of any of embodiments P91-P96, wherein the cancer is recurrent or refractory.

Examples

By way of illustration and not limitation, the following examples are presented. Compounds were named using the automated name generation tool provided in ChemBiodraw Ultra (Cambridgesoft) which generates the system name of the chemical structure with support of the Cahn-Ingold-Prelog rule of stereochemistry. Those skilled in the art can modify the procedures set forth in the illustrative embodiments to achieve the desired product.

Salts of the compounds described herein may be prepared by standard methods, such as including the acid (e.g., TFA, formic acid, or HCl) in the mobile phase during chromatographic purification, or stirring the product with a solution of the acid (e.g., aqueous HCl) after chromatographic purification.

The following abbreviations may be relevant to the present application.

Abbreviations (abbreviations)

Synthetic examples

General procedure 1 first SNAr onto pyrimidine.

To a-40 oC mixture of 5-amino-1-methylindolin-2-one (1 equivalent) in dry tetrahydrofuran [ 0.4M ] was added DIPEA (1.1 equivalent). A solution of 5-chloro-2, 4-difluoropyrimidine (1 eq.) in dry tetrahydrofuran [ 1.5M ] was slowly added to the above mixture and allowed to slowly warm to room temperature. The reaction mixture was stirred at room temperature 16 h. After this time, the reaction mixture was filtered and washed with acetonitrile. The solid was dried under vacuum to give the title compound as a tan solid.

General procedure 2 Brookfield coupling of amine to indazole CBM (Buchwald Coupling).

A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole 1a (1.1 eq), amine (1.0 eq), ruphos-Pd-G3 (0.20 eq) and NaOtBu (1.5 eq) in 1, 4-dioxane [ 0.3M ] was heated to 90 ℃ for 16 h and then cooled to room temperature. The mixture was filtered over celite and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound.

General procedure 3 methylation of amines

To a solution of amine (1.0 eq, 14.2 mmol) in DMF [ 0.15: 0.15M ] was added NaH (4.4 eq) and stirred at 0℃for 1 h. Methyl iodide (2.8 eq.) was added to the mixture at 0 ℃. The resulting reaction mixture was stirred at room temperature 16 h. The reaction mixture was then quenched with water and extracted with EA. The extracts were combined and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography eluting with PE/EA (15:1) to obtain the title compound.

General procedure 4. CBM was reduced with hydrogen.

A mixture of indazole intermediate (1.0 equivalent) and Pd/C (10 wt% palladium; 40% by weight) in EtOH: THF (1:1.5; [ 0.05M ]) was subjected to hydrogen (1 atm) at 50 ℃ for 4 h. The mixture was degassed with nitrogen and filtered through celite. The filter cake was washed sequentially with EtOH and THF. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound.

General procedure 5 BOC deprotection with HCl or TFA.

To a solution of BOC-protected amine (1.0 eq.) in 1, 4-dioxane [ 0.3M ] was added 4N HCl (14 eq.) in 1, 4-dioxane and the reaction mixture was stirred at room temperature 12 h. The volatiles were evaporated under reduced pressure to give the title compound as a solid (quantitative) which was used in the next step without further purification.

General procedure 6 SNAr of decorated TBM and CBM.

A solution of amine hydrochloride (1.0 eq), chloro/fluoropyrimidine (1.0 eq), N-diisopropylethylamine (3 to 5 eq) in DMSO [ 0.1-0.2M ] was stirred at 80℃for 2 h. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound.

Example i-1 Synthesis of intermediate 1 3- (1-methyl-6- (4- (methylamino) piperidin-1-yl) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride

Step 1 (Synthesis of tert-butyl 1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) piperidin-4-yl) (methyl) carbamate. 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (300 mg,0.6000 mmol), tert-butyl N-methyl-N- (piperidin-4-yl) carbamate (192.72 mg,0.9000 mmol), cesium carbonate (390.68 mg,1.2 mmol) and RuPhos-Pd-G3 (50.14 mg,0.0600 mmol) were added to a 1-dram vial and purged with nitrogen of 1 min. Thereafter, 1, 4-dioxane (0.8 mL) was added and the reaction mixture was stirred at 100 ℃ overnight. The product was purified from the crude mixture using column chromatography (10 g SNAP cartridge, 0-7% methanol/DCM 25 cv,7% methanol/DCM 10 CV) to give the title compound as a white solid (106 mg,0.1673 mmol,27.8% yield) ）.MS (ESI) m/z 634.0 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6 ) δ ppm 7.89 (d, J=8.19 Hz, 1 H), 7.26 - 7.51 (m, 11 H), 6.88 (d, J=1.59 Hz, 1 H), 6.82 (dd, J=9.17, 1.96 Hz, 1 H), 6.57 (d, J=8.19 Hz, 1 H), 5.44 (d, J=13.57 Hz, 4 H), 3.97 (s, 3 H), 3.87 (br d, J=12.59 Hz, 2 H), 2.77 (br t, J=11.55 Hz, 2 H), 2.70 (s, 3 H), 1.71 - 1.89 (m, 2 H), 1.61 - 1.70 (m, 2 H), 1.42 (s, 9 H).

Step 2 Synthesis of tert-butyl (1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) piperidin-4-yl) (methyl) carbamate. N- [1- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester (106 mg,0.1700 mmol) and ethanol (4.1813 mL) were dissolved in a 40 mL vial equipped with a stirring rod. The mixture was purged with nitrogen and palladium on carbon (17.8 mg,0.1700 mmol) was added. The mixture was purged again with nitrogen and then with hydrogen. The reaction was stirred under a hydrogen balloon overnight. The slurry was filtered through celite and concentrated. The residue was loaded on SNAP25G column and purified with 0-50% EtOAC/hexane (with 2% -5% MeOH additive) to give the title compound as a yellow oil (50 mg,0.110 mmol,65.6% yield).

Step 3 Synthesis of 3- (1-methyl-6- (4- (methylamino) piperidin-1-yl) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride. N- [1- [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester (50 mg,0.1100 mmol) was added to a vial equipped with a stirring bar and dichloromethane (1 mL) was added. HCl 4N (0.4400 mmol) in 1, 4-dioxane was then added to the mixture and stirred for 2 hours. The stirring bar was removed and the solvent removed in vacuo to give the title compound as an off-white solid (42 mg,0.107 mmol,97.6% yield).

Example i-2 Synthesis of intermediate 2 3- (1-methyl-6- (piperidin-4-ylamino) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride

Step 1 Synthesis of 6-bromo-3-iodo-1-methyl-indazole. To a solution of 6-bromo-1-methyl-indazole (8.00 g,37.9 mmol) in DMF (100 mL) was added NIS (25.58 g,113.7 mmol). The reaction mixture was heated to 150 ℃ for overnight and then cooled to room temperature. The volatiles were evaporated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 0-20% ethyl acetate in hexane to give the title compound as a solid (4.95 g,14.7 mmol,39% yield). MS (ESI) [ M+H ] ⁺ 336.90.

Step 2 Synthesis of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole. To a solution of 6-bromo-3-iodo-1-methyl-indazole (2.g, 5.94 mmol) in 1, 4-dioxane (30 mL) and water (3 mL) was added (2, 6-dibenzyloxy-3-pyridinyl) boronic acid (1.99 g,5.94 mmol), potassium phosphate (3.78 g,17.81 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium (ii) dichloride (0.43 g,0.5900 mmol) under N ₂, and the mixture was stirred at 80 ℃ under N ₂ for 17 h. LCMS showed complete consumption of the reaction and the desired MS was the main peak. The reaction was then cooled to room temperature and filtered. The filtrate was extracted with ethyl acetate (3×40 mL), washed with brine (2×40 mL), dried over anhydrous Na ₂SO₄ and concentrated under reduced pressure. The crude product was purified by column chromatography (silica, gradient: 0-30% ethyl acetate in petroleum ether) to the title compound as a pale yellow solid (2.1 g,4.20 mmol,71% yield) ）.MS (ES) [M+H]⁺500.3.¹H NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.91 (d, J= 8.1 Hz, 1H), 7.62 (d, J= 8.7 Hz, 1H), 7.50 – 7.24 (m, 10H), 7.12 (dd, J= 8.7, 1.4 Hz, 1H), 6.60 (d, J= 8.1 Hz, 1H), 5.45 (s, 2H), 5.43 (s, 2H), 4.05 (s, 3H).

Step 3 Synthesis of tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (4.0G, 7.99 mmol), tert-butyl 4-aminopiperidine-1-carboxylate (1.92G, 9.59 mmol), XPhos-Pd-G3 (1.35G, 1.6 mmol) and Cs ₂CO₃ (5.2G, 15.99 mmol) in 1, 4-dioxane (53.292 mL) was heated to 110℃for 28 h and then cooled to room temperature. The mixture was filtered through celite and washed with EtOAc (3×10 mL). The filtrate was concentrated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 0-90% ethyl acetate in hexane to give tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (2.81 g,4.53 mmol,57% yield) as a solid ）.MS (ESI) [M+H]⁺:620.4;¹H NMR (500 MHz, CDCl₃) δ 7.89 (d, J = 8.1 Hz, 1H), 7.47 (d, J = 8.7 Hz, 1H), 7.45 – 7.42 (m, 2H), 7.39 – 7.27 (m, 7H), 7.25 – 7.22 (m, 1H), 6.49 (d, J = 8.1 Hz, 1H), 6.36 (dd, J = 8.8, 1.9 Hz, 1H), 6.31 (d, J = 1.7 Hz, 1H), 5.46 (s, 2H), 5.38 (s, 2H), 4.06 (br, 2H), 3.98 (s, 3H), 3.73 (br, 1H), 3.57 – 3.46 (m, 2H), 3.00 (t, J = 11.9 Hz, 2H), 2.10 (dd, J = 13.0, 2.8 Hz, 2H), 1.48 (s, 9H).

Step 4 Synthesis of tert-butyl 4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate. A mixture of tert-butyl 4- [ [3- (2, 6-dibenzylox-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (600 mg,0.970 mmol) and Pearlman' S CATALYST) (167 mg,0.240 mmol) in THF (5 mL) and ethanol (3 mL) was subjected to hydrogenation 4h at 1 atm and 50 ℃. At this time, only the olefin product was observed. Additional peltier catalyst (33.4 mg,0.0500 mmol) was added and the mixture was subjected to hydrogenation 24 h at 1 atm and 50 ℃. The mixture was filtered through celite and washed with MeOH: meCN (1:1 ratio, 3 x 50.0 mL). The filtrate was concentrated under reduced pressure to give tert-butyl 4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (495 mg,0.9496 mmol,98.085% yield) as a solid ）.MS (ESI) [M+H]⁺: 442.4;¹H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.52 (dd, J = 8.8, 1.8 Hz, 1H), 6.43 (s, 1H), 5.79 (d, J = 8.2 Hz, 1H), 4.18 (dd, J = 8.7, 5.2 Hz, 1H), 3.93 – 3.86 (m, 2H), 3.81 (s, 3H), 2.96 (br, 2H), 2.60 (t, J = 7.0 Hz, 2H), 2.30 – 2.20 (m, 1H), 2.18 – 2.11 (m, 1H), 1.94 (d, J = 11.0 Hz, 2H), 1.41 (s, 9H), 1.30 – 1.19 (m, 3H).

Step 5 Synthesis of 3- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] piperidine-2, 6-dione hydrochloride. To a solution of tert-butyl 4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (1.22 g,2.76 mmol) in 1, 4-dioxane (20 mL) was added hydrogen chloride 4N (3.45 mL,13.8 mmol) in 1, 4-dioxane. The reaction mixture was stirred at 80 ℃ for 2 hours. The solid was collected by filtration to give the title compound as a white solid (1.0 g,2.65 mmol,96% yield ）.MS (ESI) m/z 342.2 [M+H]⁺.¹H NMR (400 MHz, DMSO-d6) δ ppm 10.85 (s, 1 H), 9.01 (br d, J=2.45 Hz, 1 H), 8.76 - 8.95 (m, 1 H), 7.48 (br s, 1 H), 6.72 (br s, 3 H), 4.21 - 4.30 (m, 1 H), 3.87 (s, 3 H), 3.63 - 3.75 (m, 1 H), 3.32 (br d, J=12.59 Hz, 2 H), 2.99 (br d, J=10.39 Hz, 2 H), 2.55 - 2.69 (m, 2 H), 2.22 - 2.34 (m, 1 H), 2.07 - 2.22 (m, 3 H), 1.72 (br s, 2 H).

Example i-3 Synthesis of intermediate 3- (6-amino-1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione

Step 1 Synthesis of tert-butyl 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) carbamate. 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (160 g,320 mmol) was absorbed under mechanical stirring in 1, 4-dioxane (1600 mL) in a 3000 ml multiport RBF equipped with a reflux condenser under an atmosphere of N ₂. Next, tert-butyl carbamate (56.2G, 480 mmol) was added followed by K ₂CO₃ (133G, 959 mmol) and then purged for 5 minutes, followed by XPhos Pd G2 (25.2G, 32.0 mmol), purged again for 5 minutes and then refluxed overnight at 110 ℃. The reaction mixture was filtered through celite bed, washing with ethyl acetate. The obtained filtrate was evaporated to give a crude product, which was purified using ISCO in silica gel with PE/EtOAc as eluent to give the title compound as a white solid (148 g,275 mmol,86% yield ）.MS (ESI) m/z 537.30 [M+H]⁺.¹H NMR (400 MHz, DMSO-d6) δ ppm 9.5 (s, 1H), 7.89 – 7.91 (m, 1H), 7.28 - 7.54 (m, 12 H), 6.94 (d, 1 H), 6.58 (d, 1 H), 5.41-5.45 (d, 4 H), 3.96 (s, 3 H), 1.50 (s, 9 H), 1.37 (s, 1H).

Step 2 Synthesis of tert-butyl (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) carbamate. To the flask was added tert-butyl (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) carbamate (25 g,46.6 mmol) and THF (500 mL). The mixture was purged with nitrogen 5min and then palladium on carbon (24.79 g,23.29 mmol) was added and then stirred overnight under an atmosphere of H ₂ at 55 ℃. After this time, the reaction mixture was filtered through celite, washed with THF (2L) and then the obtained filtrate was evaporated to give the title compound as a white solid (15.69 g,43.8 mmol,94% yield ）.MS (ESI) m/z 359.1 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 10.90 (s, 1H), 9.54 (s, 1H), 7.82 (m, 1H), 7.55 - 7.57 (d, 1 H), 7.04 (d, 1 H), 4.30 (m, 1 H), 3.90 (s, 3 H), 2.30 – 2.70 (m, 2 H) 2.13 – 2.37 (m, 2 H), 1.36 (s, 9H).

Step 3 Synthesis of 3- (6-amino-1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione HCl salt. Tert-butyl (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) carbamate (25 g,69.8 mmol) was dissolved in 1, 4-dioxane (250 mL) in 2L RBF with magnetic stirring. HCl (4M in dioxane) (250 ml,69.8 mmol) was added slowly and then the reaction was stirred at room temperature for 48 hours. After this time, the mixture was filtered and then the obtained solid was absorbed in methanol and then stirred sufficiently for 20 minutes, and then filtered again to give the title compound as a pale yellow solid (18 g,57.2 mmol,82% yield ）.MS (ESI) m/z 259.1 [M+H]⁺.¹H NMR (400 MHz, DMSO-d6) δ ppm 7.80 - 7.82 (d, 1 H), 7.53 (s, 1 H), 7.08 - 7.11 (d 1 H), 4.39 - 4.43 (m, 1 H), 3.99 (s, 3 H), 2.50 – 2.73 (m, 2 H), 2.38 – 2.40 (m, 1 H), 2.18 - 2.36 (m, 1 H).

Example i-4. Synthesis of intermediate 4 3- (1-methyl-6- (methyl (piperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride

Step 1 Synthesis of tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (500 mg,1 mmol), 4-aminopiperidine-1-carboxylic acid tert-butyl ester (240 mg,1.2 mmol), ruPhos-Pd-G3 (83 mg,0.1000 mmol) and Cs ₂CO₃ (651 mg,2 mmol) in 1, 4-dioxane (5 mL) was heated to 90℃for 18 h and then cooled to room temperature. The mixture was filtered through celite and washed with EtOAc (3×10 mL). The filtrate was concentrated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound as a solid （550 mg,89%）.¹H NMR (500 MHz, CDCl₃) δ 7.89 (d, J = 8.1 Hz, 1H), 7.47 (d, J = 8.7 Hz, 1H), 7.45 – 7.41 (m, 2H), 7.39 – 7.24 (m, 8H), 6.49 (d, J = 8.1 Hz, 1H), 6.36 (dd, J = 8.7, 1.9 Hz, 1H), 6.31 (d, J = 1.7 Hz, 1H), 5.46 (s, 2H), 5.38 (s, 2H), 4.13 – 4.03 (m, 2H), 3.98 (s, 3H), 3.73 (s, 1H), 3.57 – 3.50 (m, 1H), 3.00 (t, J = 12.1 Hz, 2H), 2.10 (d, J = 10.8 Hz, 2H), 1.48 (s, 9H), 1.43 – 1.35 (m, 2H).MS (ESI) [M+H]⁺ 620.5.

Step 2 Synthesis of tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] piperidine-1-carboxylate. To a solution of tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (550 mg,0.89 mmol) in DMSO (3.6 mL) and acetic acid (0.9 mL) was added an aqueous solution of formaldehyde (0.13 ml,1.8 mmol) and NaBH (OAc) ₃ (282 mg,1.3 mmol) in sequence. The reaction mixture was stirred at room temperature for 1 h. Water (10 mL) and EtOAc (25 mL) were added and the layers separated. The organic layer was washed with saturated aqueous NaHCO ₃ (mL), water (3×5 mL), brine (5 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound (467 mg,83% yield ）.¹H NMR (400 MHz, CDCl₃) δ 7.91 (d, J = 8.1 Hz, 1H), 7.56 (d, J = 9.0 Hz, 1H), 7.46 – 7.40 (m, 2H), 7.40 – 7.30 (m, 5H), 7.29 – 7.23 (m, 3H), 6.72 (dd, J = 9.3, 2.1 Hz, 1H), 6.50 (d, J = 8.1 Hz, 1H), 6.48 (d, J = 1.9 Hz, 1H), 5.47 (s, 2H), 5.38 (s, 2H), 4.25 (s, 2H), 4.01 (s, 3H), 3.80 (td, J = 10.8, 5.3 Hz, 1H), 2.84 (s, 3H), 2.83 – 2.74 (m, 2H), 1.81 – 1.64 (m, 4H), 1.49 (s, 9H).MS (ESI) [M+H]⁺ 635.5.

Step 34- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] piperidine-1-carboxylic acid tert-butyl ester. A mixture of 4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] piperidine-1-carboxylic acid tert-butyl ester (467 mg,0.74 mmol) and 20% Pd (OH) ₂/C (117 mg,25 wt%) in THF (7 mL) and EtOH (7 mL) was subjected to hydrogenation at 1 atm and 50 ℃ for 2 h. The mixture was filtered through celite. Washed with a mixture of MeCN and MeOH (1:1, 3 x 10 mL) and the filtrate concentrated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 0-20% MeOH in DCM to give the title compound as a solid （258 mg,77%）.¹H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 7.46 (d, J = 9.0 Hz, 1H), 6.87 (dd, J = 9.2, 2.0 Hz, 1H), 6.65 (d, J = 1.9 Hz, 1H), 4.23 (dd, J = 9.0, 5.1 Hz, 1H), 4.09 – 3.99 (m, 2H), 3.98 – 3.91 (m, 1H), 3.87 (s, 3H), 2.86 (br s, 2H), 2.76 (s, 3H), 2.64 – 2.56 (m, 2H), 2.33 – 2.25 (m, 1H), 2.21 – 2.13 (m, 1H), 1.67 – 1.54 (m, 4H), 1.41 (s, 9H).MS (ESI) [M+H]⁺ 456.3.

Step 4 Synthesis of 3- [ 1-methyl-6- [ methyl (4-piperidinyl) amino ] indazol-3-yl ] piperidine-2, 6-dione, hydrochloride. To a solution of tert-butyl 4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] piperidine-1-carboxylate (258 mg,0.57 mmol) in1, 4-dioxane (10 mL) was added 4M HCl (1.42 mL,5.66 mmol) in1, 4-dioxane. The reaction mixture was heated to 100 ℃ for 3 h and then cooled to room temperature. The volatiles were evaporated under reduced pressure. Et ₂ O (5 x mL) was added and the resulting precipitate was collected by filtration, washed with 1, 4-dioxane (3 x 1 mL) and Et ₂ O (10 x 2 mL) and then dried under vacuum to give the title compound （217 mg,92%）.¹H NMR (400 MHz, D₂O) δ 7.93 (d, J = 8.3 Hz, 1H), 7.65 (s, 1H), 7.31 (d, J = 8.0 Hz, 1H), 4.56 (dd, J = 10.8, 4.2 Hz, 1H), 4.18 – 4.09 (m, 1H), 4.07 (s, 3H), 3.61 (d, J = 12.6 Hz, 2H), 3.33 (s, 3H), 3.11 (t, J = 12.9 Hz, 2H), 2.91 – 2.79 (m, 2H), 2.61 – 2.48 (m, 1H), 2.44 – 2.35 (m, 1H), 2.28 (d, J = 12.0 Hz, 2H), 2.05 – 1.90 (m, 2H). as a solid annotated that no exchangeable protons were observed, containing < 1 wt% 1, 4-dioxane. MS (ESI) [ M+H ] ⁺ 356.2.2.

Example i-5 Synthesis of intermediate 5-amino-1- (2-morpholinoethyl) indol-2-one

Step 1 Synthesis of 1- (2-morpholinoethyl) -5-nitroindolin-2-one. Diisopropyl azodicarboxylate (1.091 mL,5.61 mmol) was added dropwise to a stirred solution of triphenylphosphine (1.104 g,4.21 mmol) in THF (20 mL) under nitrogen at 0 ℃ and stirred for 20 min. 2-morpholinoethyl-1-ol (736 mg,5.61 mmol) was then added and stirred 15 min. Subsequently, 5-nitroindolin-2-one (500 mg,2.81 mmol) was added and slowly warmed to 25 ℃ and stirred 16 h. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by flash column chromatography on silica gel with 60% -80% ethyl acetate/petroleum ether to give the title compound as an off-white solid (273 mg,0.937 mmol,24% yield). MS (ESI) [ M+H ] ⁺ 292.0.0.

Step 2 Synthesis of 5-amino-1- (2-morpholinoethyl) indolin-2-one. To a stirred suspension of 1- (2-morpholinoethyl) -5-nitroindol-2-one (300 mg,1.030 mmol) in ethanol (5.0 mL) and THF (5.0 mL) under nitrogen at 25 ℃ was added 10% Pd/C (150 mg). The reaction mixture was stirred under a hydrogen atmosphere for 8 h. The reaction mixture was filtered through celite and the celite pad was washed with ethanol (2×50 mL). The filtrate was concentrated under reduced pressure to obtain the title compound (280 mg,1.071 mmol,45% yield) as a gray solid. The crude product was used in the next step without further purification. MS (ESI) [ M+H ] ⁺ 262.2.

Example i-6 Synthesis of intermediate 6 5-amino-1- (4, 4-trifluorobutyl) indol-2-one

Step 1 synthesis of 5-amino-1- (4, 4-trifluorobutyl) indol-2-one the synthesis of the title compound is completed in analogy to intermediate 5 and using 4, 4-trifluorobutan-1-ol as starting material.

Example i-7 Synthesis of intermediate 7 5-amino-6-fluoro-1-methylindolin-2-one

Step 1, synthesis of 6-fluoro-1-methyl-5-nitroindoline-2-one. To a solution of 6-fluoro-1-methylindolin-2-one (100 mg,0.605 mmol) in TFA (1 mL) at 0 ℃ was added sodium nitrate (51.5 mg,0.605 mmol), and the reaction mixture was allowed to warm to room temperature and stirred for 3 hours. LCMS indicated completion of the reaction. The reaction mixture was then quenched with saturated sodium bicarbonate (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite, and concentrated. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexanes) to give the title compound as a white solid (100 mg,0.476 mmol,79% yield ）;MS (ESI) m/z 211.0 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 8.06 (br d, J=7.46 Hz, 1 H), 6.95 - 7.10 (m, 1 H), 3.63 (s, 2 H), 3.28 - 3.33 (m, 43 H), 3.24 (s, 3 H).

Step 2, synthesis of 5-amino-6-fluoro-1-methylindolin-2-one. A suspension of 6-fluoro-1-methyl-5-nitroindolin-2-one (100 mg,0.476 mmol) in methanol (10 mL) was stirred with palladium on carbon (50.6 mg,0.476 mmol) at room temperature under hydrogen (1 atm) for 2 h. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure to give the title compound (80 mg,0.444 mmol,93% yield) as an off-white solid, MS (ESI) M/z 181.2 [ M+H ] ⁺.

Example i-8 Synthesis of intermediate 8 5-amino-7-fluoro-1-methylindolin-2-one

Step 1, synthesis of 7-fluoro-1-methylindolin-2-one. To a suspension of 7-fluoroindolin-2-one (0.400 g,2.65 mmol) in water (10 mL) was added sodium hydroxide 1N (3.97 mL,3.97 mmol) and dimethyl sulfate (0.379 mL,3.97 mmol), and the reaction mixture was stirred at 120 ℃ for 40 min. LCMS indicated incomplete reaction, 20% SM remaining. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried (anhydrous sodium sulfate), filtered through celite, and concentrated. The crude product was purified by column chromatography on silica gel (0-50% ethyl acetate in hexane). The desired fraction was concentrated under reduced pressure to give the title compound 7-fluoro-1-methylindolin-2-one (0.3 g,1.82 mmol,69% yield) as a yellow solid, MS (ESI) M/z 331.2 [ M+H ] ⁺.

Step 2, synthesis of 5-amino-7-fluoro-1-methylindolin-2-one. The synthesis of the title compound was completed in analogy to intermediate 7 and using 7-fluoro-1-methylindolin-2-one as starting material.

Example i-9 Synthesis of intermediate 9 5-amino-4-fluoro-1-methylindolin-2-one

Step 1, synthesis of 4-fluoro-1-methylindolin-2-one. To a suspension of 4-fluoroindolin-2-one (1.0 g,6.62 mmol) in water (18 mL) was added sodium hydroxide 1N (9.92 mL,9.92 mmol) and dimethyl sulfate (0.695 mL,7.28 mmol) and the reaction mixture was stirred at 120 ℃ for 40 min. LCMS indicated incomplete reaction, 20% SM remaining. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried (anhydrous magnesium sulfate), filtered through celite and concentrated. The crude product was purified by column chromatography on silica gel (0-50% ethyl acetate in hexane). The desired fraction was concentrated under reduced pressure to give the title compound (1.48 g, 100%) as a white solid, MS (ESI) M/z 166.2 [ M+H ] ⁺.

Step 2, synthesis of 4-fluoro-1-methyl-5-nitroindoline-2-one. To a solution of 4-fluoro-1-methylindolin-2-one (0.68 g,4.12 mmol) in sulfuric acid (10 mL) at-30 oC was slowly added nitric acid 90% (0.195 mL,4.53 mmol) in sulfuric acid 98 (1 mL). The reaction mixture was stirred at-30 ℃ and allowed to warm to 0 ℃ over 30 min. The reaction mixture was poured onto 100 g ice and then extracted with ethyl acetate (3X 25 mL). The combined organic phases were washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous sodium sulfate, filtered through celite, and concentrated. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane) to give the title compound 4-fluoro-1-methyl-5-nitroindolin-2-one (0.260 g,1.24 mmol,30.0% yield); MS (ESI) M/z 211.2 [ M+H ] ⁺ as a yellow solid.

Step 3, synthesis of 5-amino-4-fluoro-1-methylindolin-2-one. A suspension of 4-fluoro-1-methyl-5-nitroindolin-2-one (60 mg,0.285 mmol) in methanol (10 mL) was stirred with palladium on carbon (30.4 mg,0.285 mmol) at room temperature under hydrogen (1 atm) for 2 h. The reaction mixture was filtered. The eluate was concentrated under reduced pressure to give the title compound (44 mg,0.244 mmol,86% yield) as a white solid ）;MS (ESI) m/z 181.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 6.68 (t, J = 8.56 Hz, 1H), 6.54 (d, J = 8.07 Hz, 1H), 4.76 (s, 2H), 3.52 (s, 2H), 3.04 (s, 3H).

Example i-10 Synthesis of intermediate 10 5-amino-1-isopropylindolin-2-one

Step 1 Synthesis of 1-isopropyl-5-nitroindolin-2-one A solution of 2- (2-fluoro-5-nitrophenyl) acetic acid (0.5 g,2.51 mmol), propan-2-amine (0.445 g,7.53 mmol) in DMSO (4 mL) was stirred at 60℃for 15 h. The reaction mixture was quenched with saturated aqueous ammonium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried (anhydrous sodium sulfate), filtered, and concentrated. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fraction was concentrated under reduced pressure to give 2- (2- (isopropylamino) -5-nitrophenyl) acetic acid (0.528 g, 87%) as a yellow solid. The solid 2- (2- (isopropylamino) -5-nitrophenyl) acetic acid (0.528 g) was then stirred with 10 mL of 2N HCl at room temperature for 15 hours. The resulting solid was collected by filtration to give the title compound (0.45 g,2.043 mmol,81% yield) as a yellow solid, MS (ESI) M/z 221.2 [ M+H ] ⁺.

Step 2 Synthesis of 5-amino-1-isopropylindolin-2-one A suspension of 1-isopropyl-5-nitroindolin-2-one (450 mg,2.043 mmol) in methanol (10 mL) was stirred with palladium on carbon (217 mg,2.043 mmol) at room temperature under hydrogen (1 atm) for 15 h. The reaction mixture was filtered. The eluate was concentrated under reduced pressure to give the title compound 5-amino-1-isopropylindolin-2-one (350 mg,1.840 mmol,90% yield) as an off-white solid ）;MS (ESI) m/z 191.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 6.80 (d, J=8.31 Hz, 1 H), 6.55 (d, J=1.96 Hz, 1 H), 6.43 (dd, J=8.31, 2.32 Hz, 1 H), 4.72 (br s, 2 H), 4.45 (spt, J=6.99 Hz, 1 H), 3.35 (s, 2 H), 1.34 (d, J=7.09 Hz, 6 H).

Example i-11 Synthesis of intermediate 11 5-amino-1-ethylindolin-2-one

Step 1 synthesis of 5-amino-1-ethylindolin-2-one synthesis of the title compound was accomplished in analogy to intermediate 10 and using ethylamine hydrochloride as starting material.

Example i-12 Synthesis of intermediate 12 5-amino-1- (2-hydroxyethyl) indolin-2-one

Step 1 Synthesis of 5-amino-1- (2-hydroxyethyl) indolin-2-one Synthesis of the title compound was performed in analogy to intermediate 10 and using 2-aminoethanol as starting material.

Example i-13 Synthesis of intermediate 13 5-amino-1- (2-methoxyethyl) indolin-2-one

Step 1 Synthesis of 2- (2- ((2-methoxyethyl) amino) -5-nitrophenyl) acetic acid. To a solution of 2- (2-fluoro-5-nitrophenyl) acetic acid (1,500 mg,2.51 mmol) in THF (5.0 mL) was added 2-methoxyethyl-1-amine (2,377 mg,5.02 mmol) and DIPEA (0.877 mL,5.02 mmol) under nitrogen at 25 ℃. The reaction mixture was heated to 60 ℃ and stirred 16 h. The reaction mixture was concentrated under reduced pressure to give the title compound as a yellow solid (550 mg,0.543 mmol,21% yield).

Step 2 Synthesis of 1- (2-methoxyethyl) -5-nitroindolin-2-one. To a solution of 2- (2- ((2-methoxyethyl) amino) -5-nitrophenyl) acetic acid (550 mg,2.16 mmol) in water (2.0 mL) was added 1.5N HCl (3.0 mL). The reaction mixture was heated to 60 ℃ and stirred 16 h. The reaction mixture was slowly poured into sodium bicarbonate solution at 0 ℃ and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as a light brown solid (240 mg,0.95 mmol,44% yield).

Step 3 Synthesis of 5-amino-1- (2-methoxyethyl) indolin-2-one. To a stirred solution of 1- (2-methoxyethyl) -5-nitroindolin-2-one (240 mg,0.945 mmol) in ethanol (10 mL) at 25 ℃ under nitrogen was added 10% Pd/C (101 mg). The reaction mixture was stirred under a hydrogen atmosphere for 8 h. The reaction mixture was filtered through celite and the celite pad was washed with ethanol (3×10 mL). The filtrate was concentrated under reduced pressure to obtain the title compound (200 mg,0.563 mmol,59% yield). The product was used in the next step without further purification. MS (ESI) M/z 207.2 [ M+H ] ⁺.

Example i-14 Synthesis of intermediate 14 5-amino-1-methyl-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one

Step 1 Synthesis of 3, 3-dibromo-1-methyl-5-nitro-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one. To a stirred solution of 1-methyl-5-nitro-1H-pyrrolo [2,3-b ] pyridine (800 mg,3.66 mmol) in tert-butanol (1 mL) and water (1 mL) at 0℃was added NBS (1.37 g,7.68 mmol). The reaction mixture was stirred at 25 ℃ for 2 h. The reaction mixture was monitored by TLC and LCMS. When the reaction was complete, the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (3×100 ml). The combined organic layers were evaporated to give the crude title compound, which was used without further purification.

Step 2 Synthesis of 5-amino-1-methyl-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one to a stirred solution of 3, 3-dibromo-1-methyl-5-nitro-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one (1.4 g,3.99 mmol) in AcOH (20 mL) was added zinc powder (1.30 g,20.0 mmol) and the reaction mixture was stirred at 25℃for 6H. The reaction mixture was monitored by TLC and LCMS. After this time, the reaction mixture was evaporated. The resulting residue was basified by using NaOH solution (100 ml) and extracted with 10% MeOH in DCM. The organic fraction was evaporated to give the title compound, which was used without further purification. MS (ESI) M/z 164.1 [ M+H ] ⁺.

Example i-15 Synthesis of intermediate 15 6-amino-1- (3-hydroxy-3-methylbutyl) indolin-2-one

Step 1 Synthesis of 2-methyl-4- (6-nitro-1H-indol-1-yl) butan-2-ol to a stirred solution of 6-nitro-1H-indole (1,1.0 g,6.17 mmol) in DMF (10 mL) at 25℃was added the compound 3-hydroxy-3-methylbutyl-4-methylbenzenesulfonate (2,2.30 g,8.02 mmol) and cesium carbonate (3.01 g,9.25 mmol). The reaction mixture was heated to 120 ℃ for 16 h. The reaction mixture was then cooled to room temperature and treated with water and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound, which was purified by flash column chromatography on silica with 20% -30% ethyl acetate/petroleum ether to give the title compound as a brown semi-solid (1.4 g,5.44 mmol,88% yield). MS (ESI) M/z 249.1 [ M+H ] ⁺.

Step 2 Synthesis of 3, 3-dibromo-1- (3-hydroxy-3-methylbutyl) -6-nitroindolin-2-one. To a stirred solution of 2-methyl-4- (6-nitro-1H-indol-1-yl) butan-2-ol (100 mg,0.380 mmol) in t-BuOH (1 mL) and water (1.0 mL) at 0 ℃ was added NBS (142 mg,0.797 mmol). The reaction mixture was slowly warmed to 25 ℃ and stirred 2 h. The reaction mixture was treated with water (30 mL) and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound (150 mg,94% yield). The crude product was used in the next step without further purification.

Step 3 Synthesis of 6-amino-1- (3-hydroxy-3-methylbutyl) indolin-2-one. Zinc powder (116 mg,1.78 mmol) was added to a stirred solution of 3, 3-dibromo-1- (3-hydroxy-3-methylbutyl) -6-nitroindolin-2-one (150 mg) in AcOH (3.0 mL) at 0 ℃. The reaction mixture was slowly warmed to 25 ℃ and stirred 8 h. The reaction mixture was concentrated under reduced pressure to give a crude compound. The crude product was purified by flash column chromatography on silica with 6% methanol/DCM to give the title compound as a brown gummy solid (100 mg,0.333 mmol,87% yield). MS (ESI) M/z 235.2 [ M+H ] ⁺.

Example i-16 Synthesis of intermediate 16 6-amino-1- (3-hydroxy-3-methylbutyl) -3, 3-dimethylindolin-2-one

Step 1 Synthesis of N- (2-bromo-5-nitro-phenyl) -2-methyl-prop-2-enamide. To a solution of 2-bromo-5-nitro-aniline (1.00 g,4.61 mmol) in DMA (20 mL) was added 2-methylprop-2-enoyl chloride (0.45 mL,4.61 mmol) and the reaction mixture was stirred at room temperature 12 h. Water (5.0 mL) was added and the resulting precipitate was collected by filtration, washed with water (20 mL) and then dried under vacuum to give the title compound as a solid （950 mg,72%）.¹H NMR (400 MHz, DMSO d6): δ 9.69 (s, 1H), 8.44 (s, 1H), 8.00 (d, J = 1.2 Hz, 2H), 5.97 (s, 1H), 5.63 (s, 1H), 2.00 (s, 3H).MS (ESI) [M+H]⁺ 285.1.

Step 2, synthesis of 3, 3-dimethyl-6-nitro-indolin-2-one. To a solution of N- (2-bromo-5-nitro-phenyl) -2-methyl-prop-2-enamide (950 mg,3.33 mmol) in DMF (33 mL) was added triethylamine (1.16 mL,8.33 mmol), TBAB (1.07 g,3.33 mmol) and Pd (OAc) ₂ (15.0 mg,0.07 mmol) sequentially under nitrogen and the reaction mixture was stirred at 80℃for 1 h. HCOONa (227 mg,3.33 mmol) was added and the reaction mixture was stirred at 80℃for 12 h. Water (5 mL) and ethyl acetate (15 mL) were added and the layers separated. The organic layer was washed with brine (3×10 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure. Et ₂ O (5 mL) was added and the resulting precipitate was collected by filtration and dried under vacuum to give the title compound as a solid （180 mg,26%）.¹H NMR (400 MHz, DMSO d6): δ 10.75 (s, 1H), 7.90 (dd, J = 8.2, 2.1 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.57 (d, J = 2.1 Hz, 1H), 1.30 (s, 6H).MS (ESI) [M+H]⁺ 207.0.

Step 3 Synthesis of 1- (3-hydroxy-3-methyl-butyl) -3, 3-dimethyl-6-nitro-indolin-2-one. To a solution of 3, 3-dimethyl-6-nitro-indolin-2-one (180 mg,0.87 mmol) in DMF (3 mL) was added sequentially (3-hydroxy-3-methyl-butyl) 4-methylbenzenesulfonate (271 mg,1.05 mmol) and K ₂CO₃ (362 mg,2.62 mmol), and the reaction mixture was stirred at 80 ℃ for 12 h. Water (10 mL) and ethyl acetate (60 mL) were added and the layers separated. The organic layer was washed with brine (10 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound as a solid （145 mg,57%）.¹H NMR (500 MHz, DMSO d6): δ 7.96 (dd, J = 8.1, 2.1 Hz, 1H), 7.77 (d, J = 2.1 Hz, 1H), 7.66 (d, J = 8.1 Hz, 1H), 4.50 (s, 1H), 3.86 – 3.78 (m, 2H), 1.69 – 1.62 (m, 2H), 1.31 (s, 6H), 1.17 (s, 6H).MS (ESI) [M+H]⁺275.2.

Step 4 Synthesis of 6-amino-1- (3-hydroxy-3-methyl-butyl) -3, 3-dimethyl-indolin-2-one. A mixture of 1- (3-hydroxy-3-methyl-butyl) -3, 3-dimethyl-6-nitro-indolin-2-one (145 mg,0.50 mmol) and Pd (OH) ₂/C (70.0 mg,0.10 mmol) in iPrOH (3 mL) was subjected to hydrogenation (1 atm) overnight at room temperature. The mixture was filtered through celite and washed with MeOH (10 mL). The filtrate was concentrated under reduced pressure to give the title compound (125 mg, 96%) as a solid, which was used in the next step without further purification. MS (ESI) [ M+H ] ⁺ 263.2.

Example i-17 Synthesis of intermediate 17 (2- (5-amino-2-oxoindolin-1-yl) ethyl) (methyl) carbamic acid tert-butyl ester

Step 1 synthesis of 1- (2- (methylamino) ethyl) -5-nitroindolin-2-one to a stirred solution of 2- (2-fluoro-5-nitrophenyl) acetic acid (1,1.0 g,5.02 mmol) in THF (30 mL) at 25 ℃ was added tert-butyl (2-aminoethyl) (methyl) carbamate (2,1.750 g,10.04 mmol) and DIPEA (1.754 mL,10.04 mmol) under nitrogen. The reaction mixture was heated to 65 ℃ and stirred 16 h. The reaction mixture was concentrated under reduced pressure to give a crude product as a brown liquid. The crude product was dissolved in water (20 mL) and 6.0N HCl (6 ml,36.0 mmol) was added dropwise at 25 ℃. The reaction mixture was heated to 65 ℃ and stirred 16 h. The reaction mixture was cooled to room temperature and neutralized with sodium bicarbonate and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound (400 mg,0.909 mmol,18% yield), which was used in the next step without further purification.

Step 2, synthesis of methyl (2- (5-nitro-2-oxoindolin-1-yl) ethyl) carbamic acid tert-butyl ester. DIPEA (0.397 mL,2.272 mmol) and Boc-anhydride (0.422 mL,1.818 mmol) were added dropwise to a stirred solution of 1- (2- (methylamino) ethyl) -5-nitroindol-2-one (400 mg,0.909 mmol) in DCM (10 mL) under nitrogen at 0 ℃. The reaction mixture was slowly warmed to 25 ℃ and stirred 16 h. The reaction mixture was treated with water (50 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude compound, which was purified by flash column chromatography on silica gel with 20% -40% ethyl acetate/petroleum ether to give the title compound (240 mg,0.697 mmol,77% yield).

Step 3 Synthesis of tert-butyl (2- (5-amino-2-oxoindolin-1-yl) ethyl) (methyl) carbamate. To a stirred solution of tert-butyl methyl (2- (5-nitro-2-oxoindolin-1-yl) ethyl) carbamate (240 mg,0.716 mmol) in ethanol (2.5 mL) and THF (2.5 mL) at 25 ℃ was added 10% Pd/C (120 mg) under nitrogen. The reaction mixture was stirred under a hydrogen atmosphere for 6 h. The reaction mixture was filtered through celite and the celite pad was washed with ethanol (2×50 mL). The filtrate was concentrated under reduced pressure to obtain compound 5 (200 mg,0.504 mmol,70% yield) as a dark brown liquid. The crude product was used in the next step without further purification.

Example i-18 Synthesis of intermediate 18 5-amino-1- (2, 2-difluoropropyl) indol-2-one

Step 1, synthesizing 2, 2-difluoropropyl 4-methylbenzenesulfonate. To a solution of 2, 2-difluoropropan-1-ol (1,1.0 g,10.4 mmol) in DCM (10 mL) was added triethylamine (2.18 mL,15.61 mmol) at 25 ℃. The mixture was cooled to 0 ℃ and 4-dimethylaminopyridine (0.127 g,1.041 mmol) was added followed by 4-methylbenzenesulfonyl chloride (2.381 g,12.49 mmol). The reaction mixture was slowly warmed to 25 ℃ and stirred 16 h. The resulting reaction mixture was added to water (25 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as an off-white solid (2.0 g,7.95 mmol,76% yield). The crude product was used in the next step without further purification. MS (ESI) M/z 268.0 [ M+H ₂ O ].

Step 2 Synthesis of 1- (2, 2-difluoropropyl) -5-nitro-1H-indole. To a solution of 2, 2-difluoropropyl 4-methylbenzenesulfonate (931 mg,3.70 mmol) in DMF (5 mL) was added cesium carbonate (1507 mg,4.63 mmol) and 5-nitro-1H-indole (3,500.0 mg,3.08 mmol) at 25 ℃. The mixture was heated to 120 ℃ and stirred 16 h. The reaction mixture was quenched by addition of water (15 mL) and extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by flash column chromatography on silica with 35% -40% ethyl acetate/petroleum ether to give the title compound as a pale yellow solid (250.0 mg,0.997 mmol,32% yield). MS (ESI) M/z 240.9 [ M+H ] ⁺.

Step 3 Synthesis of 3, 3-dibromo-1- (2, 2-difluoropropyl) -5-nitroindolin-2-one. To a solution of 1- (2, 2-difluoropropyl) -5-nitro-1H-indole (250.0 mg,0.997 mmol) in t-butanol (2.5 ml,26.1 mmol) and water (2.5 mL) at 0 ℃ was added NBS (373 mg,2.094 mmol) in portions. The reaction mixture was slowly warmed to 25 ℃ and stirred 6 h. The reaction mixture was quenched with water (15 mL) and extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as a pale yellow solid (350.0 mg,0.619 mmol,62% yield). MS (ESI) M/z 415.0 [ M+H ] ⁺.

Step 4 Synthesis of 5-amino-1- (2, 2-difluoropropyl) indolin-2-one. Zinc powder (202 mg,3.10 mmol) was slowly added to a stirred solution of 3, 3-dibromo-1- (2, 2-difluoropropyl) -5-nitroindolin-2-one (350.0 mg,0.619 mmol) in acetic acid (5.0 mL) at 25℃and stirring was continued for 6 h. The reaction mixture was concentrated under reduced pressure and diluted by the addition of water (15 mL). The resulting residue was extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as a brown solid (150.0 mg,0.299 mmol,48% yield). The crude product was used in the next step without further purification. MS (ESI) M/z 227.0 [ M+H ] ⁺.

Example i-19 Synthesis of intermediate 19 5-amino-1- (3-fluoro-3-methylbutyl) indolin-2-one

Step 1 Synthesis of 5-amino-1- (3-fluoro-3-methylbutyl) indolin-2-one. The title compound was synthesized in analogy to intermediate 18, using 3-fluoro-3-methylbutan-1-ol as starting material.

Example i-20 Synthesis of intermediate 20 5-amino-1- (2-fluoro-2-methylpropyl) indolin-2-one

Step 1, synthesizing 2-fluoro-2-methylpropyl trifluoro methane sulfonate. To a stirred mixture of trifluoromethanesulfonic anhydride (2.75 mL,16.28 mmol) in DCM (10 mL) at-10 ℃ was slowly added 2, 6-lutidine (2.26 mL,19.5 mmol) and then a solution of 2-fluoro-2-methylpropan-1-ol (1.50 g,16.3 mmol) in DCM (35 mL) was added dropwise. The reaction mixture was stirred at 0 ℃ for 4 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was dissolved in DCM and washed with 1.0N HCl solution, followed by saturated bicarbonate solution and then brine. The organic phase was dried over sodium sulfate, filtered and concentrated to give the title compound as a brown liquid (2.5 g,11.2 mmol,68% yield). The crude product was used in the next step without purification.

Step 2 Synthesis of 1- (2-fluoro-2-methylpropyl) -5-nitro-1H-indole. To a stirred solution of 5-nitro-1H-indole (700 mg,4.32 mmol) in DMF (10 mL) was added the compound 2-fluoro-2-methylpropyl triflate (2419 mg 10.79 mmol). The reaction mixture was heated to 100 ℃ and stirred 4 h. The reaction mixture was poured into ice-cold water and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated to give the title compound (600 mg,2.41 mmol,56% yield). The crude product was used in the next step without purification. MS (ESI) M/z 237.1 [ M+H ] ⁺.

Synthesis of 5-amino-1- (2-fluoro-2-methylpropyl) indolin-2-one. To a stirred solution of 1- (2-fluoro-2-methylpropyl) -5-nitro-1H-indole (100 mg,0.423 mmol) in t-butanol (2.0 mL) at 0 ℃ was added bromine (0.218 mL,4.23 mmol) and the reaction mixture was stirred 5 min. Water (2 mL) was then slowly added and warmed to 25 ℃ and stirred 4 h. The reaction mixture was treated with water and extracted with ethyl acetate (3×10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 110 mg crude material. The residue was dissolved in acetic acid (3.5 mL) and zinc powder (88 mg,1.34 mmol) was added and stirred at 25 ℃ for 1 h. The reaction mixture was concentrated under reduced pressure to give a crude product. The crude product was dissolved in ethyl acetate and treated with sodium bicarbonate solution. The organic layer was dried over sodium sulfate, filtered and concentrated to give the title compound (55 mg,0.132 mmol,49% yield). The crude product was used in the next step without purification.

Example i-21 Synthesis of intermediate 21 5-amino-1- (2, 2-trifluoroethyl) indolin-2-one

Step 1 Synthesis of 5-nitro-1- (2, 2-trifluoroethyl) indoline-2, 3-dione. To a solution of 5-nitroindoline-2, 3-dione (0.96 g,5.00 mmol) in DMF (35.0 mL) cooled to 0 ℃ was added NaH (60% dispersion in mineral oil, 220.0 mg,5.50 mmol). After stirring 45 min at 0 ℃, 2-trifluoroethyl triflate (1.28 g,5.50 mmol) was added. The reaction mixture was warmed to room temperature for 1h a. Saturated aqueous NH ₄ Cl (300.0 mL) and EtOAc were added and the layers separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with water, brine, dried (Na ₂SO₄), filtered and concentrated under reduced pressure. The material was purified by column chromatography on silica gel using a gradient of 20% -70% ethyl acetate in hexane to give the title compound as a solid （910.0 mg,66%）.¹H NMR (500 MHz, DMSO) δ 8.60 (dd, J = 8.8, 2.5 Hz, 1H), 8.30 (d, J = 2.4 Hz, 1H), 7.55 (d, J = 8.8 Hz, 1H), 4.77 (q, J = 9.3 Hz, 2H).

Step 2 Synthesis of 5-amino-1- (2, 2-trifluoroethyl) indoline-2, 3-dione. A mixture of 5-nitro-1- (2, 2-trifluoroethyl) indoline-2, 3-dione (548..0 mg,2.00 mmol) and 10% Pd/C (106.0 mg,0.10 mmol) in methanol (25.0 mL) was shaken in a Parr flask (PARR FLASK) under 50 psi hydrogen atmosphere at room temperature for 1 h. The mixture was filtered through celite and washed with MeOH. The filtrate was concentrated under reduced pressure to give the title compound (450.0 mg, 92%) as a solid, which was used in the next step without further purification. MS (ESI) [ M+H ] ⁺ 247.1.1.

Step 3 Synthesis of 5-amino-1- (2, 2-trifluoroethyl) indolin-2-one. A mixture of 5-amino-1- (2, 2-trifluoroethyl) indoline-2, 3-dione (450.0 mg,1.84 mmol) and hydrazine hydrate (65%, 10.0 mL,133.0 mmol) was heated to 115 ℃ for 1h and then cooled to room temperature. The volatiles were evaporated under reduced pressure. The material was purified by reverse phase chromatography (C18) using a gradient of 10% -70% acetonitrile and water (ammonium formate buffer pH 4) to give the title compound as a solid （150.0 mg,35%）.¹H NMR (500 MHz, DMSO) δ 6.79 (d, J = 8.3 Hz, 1H), 6.57 (s, 1H), 6.47 (dd, J = 8.3, 2.1 Hz, 1H), 4.81 (s, 2H), 4.47 (q, J = 9.5 Hz, 2H), 3.54 (s, 2H).MS (ESI) [M+H]⁺ 231.1.

Example i-22 Synthesis of intermediate 22 4-amino-5-methylcycloheptane-1-carboxylic acid benzyl ester

Step 1 Synthesis of 1-benzyl 4-ethyl 5-oxoazepane-1, 4-dicarboxylic acid ester BF ₃OEt₂ (1.358 mL,10.72 mmol) was added to a stirred solution of benzyl 4-oxopiperidine-1-carboxylate (2.5 g,10.72 mmol) and ethyl 2-diazonoacetate (9.78 g,12.86 mmol) in THF (50 mL) at-25 ℃. The reaction mixture was stirred at 25 ℃ for 4 h. After that, the reaction mixture was basified with sodium bicarbonate solution (100 ml) and the organics were extracted with EtOAc (3×100 ml). The combined organic fractions were dried and concentrated to give crude material. It was purified by Combi-flash column chromatography, in which the product eluted at 15% EA in PE. The product containing fractions were combined and concentrated to give 1-benzyl 4-ethyl 5-oxo-azepane-1, 4-dicarboxylic acid ester (3.5 g,3.40 mmol,31.7% yield) as a colourless liquid. (ESI) [ m+h ] ⁺ 320.0, rt=2.15.

Step 2 Synthesis of 1-benzyl 4-ethyl 4-methyl-5-oxoazepane-1, 4-dicarboxylic acid ester Cs ₂CO₃ (7.14 g,21.92 mmol) and MeI (1.371 mL,21.92 mmol) were added to a stirred solution of 1-benzyl 4-ethyl 5-oxoazepane-1, 4-dicarboxylic acid ester (3.5 g,10.96 mmol) in DMF (8 mL) at 0 ℃. The reaction mixture was warmed to room temperature and stirred at 25 ℃ for 6 h. The reaction mixture was evaporated to obtain a crude material, which was purified by combi-flash column chromatography. The product eluted at 25% -30% EA in PE and the fractions containing the product are combined and concentrated to give 1-benzyl 4-ethyl 4-methyl-5-oxoazepane-1, 4-dicarboxylic acid ester as a colourless liquid （2.5 g,76%）.¹H NMR (400 MHz, DMSO) δ 7.35 (br m, 5H), 5.07 (br m, 2H), 4.15 ( br m, 2H), 3.68 (m, 2H), 3.40 (br m, 1H), 2.81 (br m, 2H), 2.15 (br m, 1H), 1.59 (br m, 1H), 1.19 – 1.24 (br m, 6H).MS (ESI) [M+H]⁺ 334.1

Step 3 Synthesis of benzyl 4-methyl-5-oxoazepane-1-carboxylate to a stirred solution of 1-benzyl 4-ethyl 4-methyl-5-oxoazepane-1, 4-dicarboxylic acid ester (2.5 g,5.70 mmol) in MeOH (20 mL) and water (20 mL) was added KOH (0.959 g,17.10 mmol) at 25 ℃. The reaction mixture was stirred at 60 ℃ for 4 h. Thereafter, the reaction mixture was diluted with water (50 ml) and extracted with DCM (2×100 ml). It was purified via reverse phase prep HPLC with a gradient of 2% -98% MeCN in 5mm ammonium formate pH 3.3. Fractions containing the desired product were combined and concentrated to give benzyl 4-methyl-5-oxo-azepane-1-carboxylate as a yellow oil （1.50 g,80%）.¹H NMR (400 MHz, DMSO) δ 7.31 - 7.39 (m, 5H), 5.07 (s, 2H), 3.82 - 3.98 (m, 2H), 3.46 (m, 1H), 3.34 (m, 1H), 3.19 (m, 1H), 3.01 (m, 1H). 2.59 (m, 1H), 1.66 (m, 1H), 1.30 (m, 1H), 0.95 (d, 2H).MS (ESI) [M+H]⁺ 262.2

Step 4 Synthesis of benzyl 4-amino-5-methylazepan-1-carboxylate to a stirred solution of benzyl 4-methyl-5-oxoazepan-1-carboxylate (1.0 g,3.02 mmol) in 2-propanol (20 mL) was added ammonium acetate (4.66 g,60.5 mmol) and NaCNBH ₄ (1.140 g,18.14 mmol) at 25 ℃. The reaction mixture was stirred at 50 ℃ for 3h and monitored by TLC and LCMS. Thereafter, the reaction mixture was diluted with water (80 ml) and extracted with DCM (3×80 ml). The organic fraction was concentrated to give benzyl 4-amino-5-methylazepan-1-carboxylate (700 mg, 65%) as a yellow solid. MS (ESI) [ m+h ] ⁺ 263.2, rt=0.78

Example i-23 Synthesis of intermediate 23 4-amino-3-methylazepan-1-carboxylic acid benzyl ester

Step 1 Synthesis of 1-benzyl 3-ethyl 4-oxoazepane-1, 3-dicarboxylic acid ester BF ₃OEt₂ (2.72 mL,21.43 mmol) was added to a stirred solution of benzyl 3-oxopiperidine-1-carboxylate (5.0 g,21.43 mmol) and ethyl 2-diazonoacetate (21.20 g,27.9 mmol) in THF (50 mL) at-78 ℃. The reaction mixture was warmed to room temperature and stirred at 25 ℃ for 4 h. After that, the reaction mixture was basified with sodium bicarbonate solution (100 ml) and the organics were extracted with EtOAc (3×100 ml). The combined organic fractions were dried and concentrated to give crude material. It was purified by Combi-flash column chromatography, in which the product eluted at 20% -30% EA in PE. The product containing fractions were combined and concentrated to give 1-benzyl 3-ethyl 4-oxo-azepane-1, 3-dicarboxylic acid ester (4.5 g,10.00 mmol,46.7% yield) as a colourless liquid. (ESI) [ m+h ] ⁺ 320.1.1, rt=2.11.

Step 2 Synthesis of 1-benzyl 3-ethyl 3-methyl-4-oxoazepane-1, 3-dicarboxylic acid ester Cs ₂CO₃ (6.53 g,20.04 mmol) and MeI (1.253 mL,20.04 mmol) were added to a stirred solution of 1-benzyl 3-ethyl 4-oxoazepane-1, 3-dicarboxylic acid ester (4.0 g,10.02 mmol) in DMF (1 mL) at 0 ℃. And the reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was warmed to room temperature and stirred at 25 ℃ for 6 h. The reaction mixture was evaporated to obtain a crude material, which was purified by combi-flash column chromatography. The product eluted under 20% EtOAc in PE and the fractions containing the product were combined and concentrated to give 1-benzyl 3-ethyl 3-methyl-4-oxoazepane-1, 3-dicarboxylic acid ester (2.2 g,5.61 mmol,56.0% yield) as a colorless liquid ）.¹H NMR (400 MHz, DMSO) δ 7.32 - 7.40 (m, 5H), 5.08 (s, 2H), 3.34 – 4.04 (m, 6H), 2.71 (m, 2H), 1.64 – 1.78 (m, 3H). 1.10 - 1.24 (m, 5H).MS (ESI) [M+H]⁺ 334.2

Step 3 Synthesis of benzyl 3-methyl-4-oxoazepane-1-carboxylate to a stirred solution of 1-benzyl 3-ethyl 3-methyl-4-oxoazepane-1, 3-dicarboxylic acid ester (1.0 g,2.55 mmol) in MeOH (7.5 mL) and H ₂ O (7.5 mL) was added KOH (0.429 g,7.65 mmol) and the reaction mixture stirred at 60℃for 2H. The reaction mixture was monitored by TLC and LCMS. The reaction mixture was stirred at 60 ℃ for 4 h. Thereafter, the reaction mixture was diluted with water (50 ml) and extracted with DCM (2×100 ml). It was purified via reverse phase prep HPLC with a gradient of 2% -98% MeCN in 5 mm ammonium formate pH 3.3. Fractions containing the desired product were combined and concentrated to give benzyl 3-methyl-4-oxo-azepane-1-carboxylate as a yellow oil （550 mg,76%）.¹H NMR (400 MHz, DMSO) δ 7.29 - 7.37 (m, 5H), 5.08 (s, 2H), 3.33 – 4.04 (m, 6H), 2.51 (m, 2H), 1.99 (m, 1H), 1.76 (m, 1H), 1.64 (m, 1H). 1.09 - 1.23 (m, 6H).MS (ESI) [M+H]⁺ 262.1

Step 4 Synthesis of benzyl 4-amino-3-methylazepan-1-carboxylate to a stirred solution of benzyl 3-methyl-4-oxoazepan-1-carboxylate (100 mg,0.352 mmol) in 2-propanol (4 mL) was added ammonium acetate (543 mg,7.04 mmol) and NaCNBH ₄ (133 mg,2.112 mmol) at 25 ℃. The reaction mixture was stirred at 50 ℃ for 3 h. After this time, the reaction mixture was diluted with water and extracted with EtOAC. The organic fraction was concentrated to give benzyl 4-amino-3-methylazepan-1-carboxylate (100 mg, 40%) as a yellow solid. MS (ESI) [ m+h ] ⁺ 263.2, rt=1.35.

EXAMPLE S1.3 Synthesis of 6- ((5- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) -5-azaspiro [3.5] non-8-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (1)

Step 1 Synthesis of 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one. To a solution of 5-amino-1-methylindolin-2-one (2.0 g,12.3 mmol) in tetrahydrofuran (32 mL) was added DIPEA (2.37 ml,13.6 mmol) at-40 ℃. A solution of 5-chloro-2, 4-difluoropyrimidine (1.86 g,12.3 mmol) in dry tetrahydrofuran (8 mL) was slowly added to the above mixture and allowed to slowly warm to room temperature. The reaction mixture was stirred at room temperature 16 h. After this time, the reaction mixture was filtered and washed with acetonitrile (about 50 mL). The solid was dried under vacuum to give the title compound as a tan solid. MS (ESI) [ M+H ] ⁺ 293.1.

Step 2 Synthesis of tert-butyl 8- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] -5-azaspiro [3.5] nonane-5-carboxylate. To a solution of 3- (6-amino-1-methyl-indazol-3-yl) piperidine-2, 6-dione hydrochloride (200 mg,0.68 mmol), 8-oxo-5-azaspiro [3.5] nonane-5-carboxylic acid tert-butyl ester (201 mg,0.81 mmol) and AcOH (1 mL) in DMSO (5 mL) was added decaborane (38 mg,0.34 mmol). The reaction mixture was stirred at room temperature 5 h and the volatiles were evaporated under reduced pressure. The residue was purified by reverse phase chromatography (C18) using a gradient of 0-80% MeCN and 10 mM ammonium formate in water to give the title compound as a solid （325 mg,76%）.MS (ESI) [M+H]⁺ 482.3.¹H NMR (400 MHz, DMSO-d₆) δ 10.82 (s, 1H), 7.32 (d, J = 8.7 Hz, 1H), 6.52 – 6.40 (m, 1H), 6.48 (br s, 1H), 5.78 (d, J = 8.7 Hz, 1H), 4.18 (dd, J = 8.8, 5.2 Hz, 1H), 3.82 (s, 3H), 3.73 – 3.68 (m, 2H), 2.75 – 2.68 (m, 1H), 2.62 – 2.59 (m, 2H), 2.30 – 2.13 (m, 4H), 1.98 – 1.86 (m, 4H), 1.72 – 1.60 (m, 3H), 1.40 (s, 9H), 1.06 – 0.98 (m, 1H).

Step 3 Synthesis of 3- [6- (5-azaspiro [3.5] non-8-ylamino) -1-methyl-indazol-3-yl ] piperidine-2, 6-dione 2, 2-trifluoro-acetic acid. To a solution of tert-butyl 8- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] -5-azaspiro [3.5] nonane-5-carboxylate (325 mg,0.67 mmol) in DCM (5 mL) was added trifluoroacetic acid (0.31 ml,4.1 mmol), and the reaction mixture was stirred at room temperature 18 h. The volatiles were evaporated under reduced pressure to give the title compound as a solid, which was used in the next step without further purification .MS (ESI) [M+H]⁺ 382.3.¹H NMR (500 MHz, DMSO-d6) δ 10.82 (s, 1H), 8.98 (d, J = 8.5 Hz, 1H), 8.75 – 8.71 (m, 1H), 7.37 (d, J = 8.7 Hz, 1H), 6.55 (dd, J = 8.7, 1.8 Hz, 1H), 6.51 (br s, 1H), 4.19 (dd, J = 8.9, 5.1 Hz, 1H), 3.83 (s, 3H), 3.63 – 3.57 (m, 1H), 3.30 – 3.27 (m, 1H), 3.02 – 2.96 (m, 1H), 2.63 – 2.60 (m, 2H), 2.36 (d, J = 12.9 Hz, 1H), 2.30 – 2.23 (m, 4H), 2.17 – 2.12 (m, 2H), 2.01 – 1.97 (m, 1H), 1.95 – 1.87 (m, 2H), 1.63 – 1.58 (m, 1H), 1.40 – 1.32 (m, 1H).

Step 4 Synthesis of 3- [6- [ [5- [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -5-azaspiro [3.5] non-8-yl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione. To a solution of 3- [6- (5-azaspiro [3.5] non-8-ylamino) -1-methyl-indazol-3-yl ] piperidine-2, 6-dione 2, 2-trifluoroacetic acid (20 mg, 40. Mu. Mol) in NMP (0.7 mL) was added 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (11 mg, 40. Mu. Mol) under N ₂ (1 atm). The reaction mixture was heated to 160 ℃ for 18 h and then cooled to room temperature. The crude reaction mixture was purified by reverse phase chromatography (C18) using a gradient of 0-100% MeCN and water (0.1% formic acid) to give the title compound (13 mg, 44%) as a solid. LCMS-C ₃₄H₃₆ClN₉O₃ theory of 653.3, found 654.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.62 (s, 1H), 8.01 (s, 1H), 7.39 (s, 1H), 7.38 – 7.36 (m, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 6.48 – 6.45 (m, 2H), 5.65 (d, J = 8.8 Hz, 1H), 4.25 (d, J = 12.1 Hz, 1H), 4.17 (dd, J = 8.9, 5.1 Hz, 1H), 3.83 –3.80 (m, 1H), 3.81 (s, 3H) 3.51 – 3.50 (m, 2H), 3.09 (s, 3H), 2.88 (t, J = 10.0 Hz, 1H), 2.62 – 2.59 (m, 2H), 2.38 –2.34 (m, 2H), 2.26 – 2.24 (m, 1H), 2.19 – 2.12 (m, 2H), 1.96 –1.88 (m, 3H), 1.67 – 1.56 (m, 3H), 1.07 – 0.99 (m, 1H).

EXAMPLE S2.3 Synthesis of 3- (6- (8- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -5-azaspiro [3.5] non-5-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (2)

Step 1 Synthesis of 2- [ 2-bis- [3, 5-bis- (trifluoromethyl) phenyl ] phosphanyl-3, 6-dimethoxy-phenyl ] -N1, N1, N3, N3-tetramethyl-benzene-1, 3-diamine [2- [2- (methylamino) phenyl ] -methylsulfonyloxy-palladium. A mixture of palladium (II) 2 '-methylamino-1, 1' -biphenyl-2-yl) methanesulfonate dimer (406 mg,0.53 mmol) and 2- [ 2-bis- [3, 5-bis- (trifluoromethyl) phenyl ] phosphanyl-3, 6-dimethoxy-phenyl ] -N1, N1, N3, N3-tetramethyl-benzene-1, 3-diamine (800 mg,1.06 mmol) was evacuated and backfilled with argon. DCM (5 mL) was added via syringe and the mixture was stirred at room temperature 2 h. The reaction mixture was diluted with Et ₂ O (5 mL) and filtered. The filtrate was concentrated and pentane (5 mL) was added to the residue and the mixture was concentrated under reduced pressure to give the title compound as a yellow solid （1.08 g,90%）.¹H NMR (500 MHz, MeOD-d₄) δ 8.38 (s, 1H), 8.13 (s, 1H), 8.00 (t, J = 8.1 Hz, 1H), 7.74 (d, J = 11.2 Hz, 2H), 7.62 (d, J = 11.4 Hz, 2H), 7.59 – 7.55 (m, 1H), 7.51 (d, J = 9.0 Hz, 1H), 7.42 (dd, J = 7.5, 1.4 Hz, 1H), 7.34 – 7.28 (m, 3H), 7.25 (t, J = 7.3 Hz, 1H), 7.21 (d, J = 8.1 Hz, 1H), 7.16 – 7.13 (m, 1H), 7.03 (d, J = 8.1 Hz, 1H), 6.84 (t, J = 7.7 Hz, 1H), 6.33 – 6.29 (m, 1H), 3.71 (s, 3H), 3.42 (s, 3H), 2.78 (s, 6H), 2.69 (s, 3H), 2.61 – 2.54 (m, 1H), 2.15 (dd, J = 5.9, 2.9 Hz, 3H), 2.09 (s, 6H).¹⁹F NMR (471 MHz, MeOD-d₄) δ –64.20, –64.42.³¹P NMR (203 MHz, MeOD-d₄) δ 33.80.

Step 2 Synthesis of tert-butyl N- [5- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] carbamate. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (600 mg,1.20 mmol), tert-butyl N- (5-azaspiro [3.5] non-8-yl) carbamate (375 mg,1.56 mmol), naOtBu (288 mg,3.0 mmol), 2- [ 2-bis- [3, 5-bis- (trifluoromethyl) phenyl ] phosphanyl-3, 6-dimethoxy-phenyl ] -N1, N1, N3, N3-tetramethyl-benzene-1, 3-diamine [2- [2- (methylamino) phenyl ] -methylsulfonyloxy-palladium (137 mg,0.12 mmol) in cyclopentylmethyl ether (12 mL) was heated to 85℃for 4h and then cooled to room temperature. The mixture was filtered through celite and washed with EtOAc (3×15 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-60% EtOAc in hexanes to give the title compound as a solid （600 mg,76%）.MS (ESI) [M+H]⁺ 660.2.¹H NMR (400 MHz, DMSO-d6) δ 7.88 (d, J = 8.1 Hz, 1H), 7.50 – 7.43 (m, 3H), 7.42 – 7.32 (m, 5H), 7.32 – 7.25 (m, 3H), 6.79 (d, J = 7.5 Hz, 1H), 6.73 (d, J = 1.5 Hz, 1H), 6.62 (dd, J = 9.0, 1.8 Hz, 1H), 6.57 (d, J = 8.1 Hz, 1H), 5.44 (s, 2H), 5.42 (s, 2H), 3.95 (s, 3H), 3.69 – 3.57 (m, 1H), 3.51 – 3.43 (m, 1H), 3.12 – 3.00 (m, 1H), 2.17 – 2.04 (m, 4H), 1.98 – 1.89 (m, 1H), 1.80 – 1.68 (m, 2H), 1.59 – 1.48 (m, 2H), 1.38 (s, 9H), 1.27 – 1.16 (m, 1H).

Step 3:N- [5- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] -N-methyl-carbamic acid tert-butyl ester. To an ice-cold solution of N- [5- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] carbamic acid tert-butyl ester (1.05 g,1.59 mmol) and NaH (159 mg,3.98 mmol, 60% dispersion in mineral oil) in DMF (6 mL) was added methyl iodide (198 μl,3.18 mmol) and the mixture was stirred at room temperature for 2 h. Another batch was prepared using the same procedure. Water (50 mL) was added and the combined mixture was extracted with EtOAc (3X 75 mL). The organic fraction was washed with brine, dried over Na ₂SO₄, filtered and concentrated. The residue was purified by reverse phase chromatography (C18) using a gradient of 20% -100% MeCN and 10mM ammonium formate in water to give the title compound as a solid (700 mg,65% combined yield). MS (ESI) [ M+H ] ⁺ 674.4.674.

Step 4 Synthesis of N- [5- [3- (2, 6-dioxo-3-piperidyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] -N-methyl-carbamic acid tert-butyl ester. A mixture of N- [5- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] -N-methyl-carbamic acid tert-butyl ester (700 mg,1.04 mmol) and Peltier's catalyst (365 mg,0.26 mmol) in EtOH (15 mL) and THF (15 mL) was stirred under a hydrogen atmosphere (1 atm) at 50℃for 2h. The mixture was filtered through celite, washed with MeOH (3×15 mL) and the filtrate was concentrated to give the title compound as a solid (425 mg, 83%) which was used in the next step without further purification .MS (ESI) [M+H]⁺ 496.2.¹H NMR (400 MHz, DMSO-d₆) δ 10.86 (s, 1H), 7.48 (d, J = 8.7 Hz, 1H), 6.77 (s, 1H), 6.77 – 6.73 (m, 1H), 4.30 – 4.22 (m, 1H), 3.89 (s, 3H), 3.62 – 3.53 (m, 1H), 3.16 – 3.05 (m, 1H), 2.70 – 2.57 (m, 5H), 2.38 – 2.25 (m, 1H), 2.19 – 2.07 (m, 4H), 2.06 – 1.93 (m, 2H), 1.93 – 1.81 (m, 1H), 1.78 – 1.67 (m, 1H), 1.62 – 1.31 (m, 4H), 1.41 (s, 9H).

Step 5 Synthesis of 3- [ 1-methyl-6- [8- (methylamino) -5-azaspiro [3.5] non-5-yl ] indazol-3-yl ] piperidine-2, 6-dione hydrochloride. To a solution of N- [5- [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] -5-azaspiro [3.5] non-8-yl ] -N-methyl-carbamic acid tert-butyl ester (425 mg,0.86 mmol) in DCM (4 mL) was added 4N HCl in 1, 4-dioxane (4 ml,16 mmol), and the mixture was stirred at room temperature 4 h. The resulting precipitate was collected by filtration, washed with Et ₂ O (2 x 5 mL), and dried under vacuum to give the title compound (340 mg, 92%) as a solid, which was used in the next step without further purification .MS (ESI) [M+H]⁺ 396.3.¹H NMR (500 MHz, DMSO-d₆) δ 10.49 (s, 1H), 9.08 (br s, 2H), 7.56 (d, J = 8.8 Hz, 1H), 6.95 (s, 1H), 6.87 (dd, J = 8.8, 1.4 Hz, 1H), 4.27 (dd, J = 8.8, 5.2 Hz, 1H), 3.92 (s, 3H), 3.60 (dt, J = 13.6, 3.2 Hz, 1H), 3.43 – 3.32 (m, 1H), 3.25 – 3.16 (m, 1H), 2.68 – 2.63 (m, 2H), 2.56 (t, J = 5.3 Hz, 3H), 2.49 – 2.43 (m, 1H), 2.39 – 2.29 (m, 1H), 2.27 – 2.20 (m, 2H), 2.20 – 2.13 (m, 2H), 2.13 – 2.04 (m, 2H), 1.96 – 1.89 (m, 1H), 1.82 – 1.70 (m, 1H), 1.64 – 1.53 (m, 2H).

Step 6 Synthesis of 3- [6- [8- [ [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -methyl-amino ] -5-azaspiro [3.5] non-5-yl ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione. To a mixture of 3- [ 1-methyl-6- [8- (methylamino) -5-azaspiro [3.5] non-5-yl ] indazol-3-yl ] piperidine-2, 6-dione hydrochloride (46.5 mg,0.11 mmol) and DIPEA (110 μl,0.61 mmol) in DMSO (1 mL) was added 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (30 mg,0.10 mmol) at room temperature, and the reaction mixture was heated to 80 ℃ for 16 h. The crude reaction mixture was purified by preparative HPLC (BEH column, C18) using a gradient of 48% -58% MeCN and 10 mM ammonium formate in water to give the title compound (14.9 mg, 22%) as a solid. LCMS: C ₃₄H₃₆ClN₉O₃ theory 667.3, found 668.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.49 (br s, 1H), 8.29 (s, 1H), 8.00 (s, 1H), 7.56 (s, 1H), 7.54 – 7.49 (m, 2H), 6.93 (d, J = 8.3 Hz, 1H), 6.80 – 6.76 (m, 2H), 4.89 – 4.80 (m, 1H), 4.26 (dd, J = 8.6, 5.2 Hz, 1H), 3.91 (s, 3H), 3.57 (d, J = 13.3 Hz, 1H), 3.54 (s, 2H), 3.15 (s, 3H), 2.89 (s, 3H), 2.74 – 2.60 (m, 2H), 2.40 – 2.30 (m, 1H), 2.27 – 2.16 (m, 2H), 2.13 – 2.05 (m, 2H), 2.01 – 1.88 (m, 3H), 1.69 – 1.55 (m, 2H), 1.55 – 1.41 (m, 2H). notes that one CH is masked by the water signal.

EXAMPLE S3.Synthesis of 3- (6- (((trans) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) cyclohexyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (3)

The title compound was synthesized in analogy to example S2, using trans-t-butyl N- (4-aminocyclohexyl) carbamate as starting material. The title compound was isolated as an off-white solid (36.7 mg,0.05 mmol,20.6% yield). LCMS C ₃₃H₃₆ClN₉O₃ theory 641, found 642 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 8.14 (s, 1H), 7.60 (s, 1H), 7.54 (d, J = 8.2 Hz, 1H), 7.42 (d, J = 8.7 Hz, 1H), 7.00 (d, J = 8.4 Hz, 1H), 6.60 (d, J = 8.9 Hz, 1H), 6.55 (s, 1H), 4.30 (s, 1H), 4.22 (dd, J = 9.0, 5.1 Hz, 1H), 3.86(s,3H), 3.59 (s, 2H), 3.36 (s, 1H), 3.15 (s, 3H), 2.95 (s, 3H), 2.62 (s, 2H), 2.28 (s, 1H), 2.20 – 2.09 (m, 3H), 1.76 (d, J = 11.8 Hz, 1H), 1.70 (s, 3H), 1.25 (s, 2H).

EXAMPLE S4.3 Synthesis of 3- (6- ((2R, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methylpiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (4)

Step 1 Synthesis of N- [ (2R, 4R) -1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -2-methyl-4-piperidinyl ] carbamate. The title compound was synthesized according to general procedure 2 using tert-butyl N- [ (2 r,4 r) -2-methyl-4-piperidinyl ] carbamate as reactant in toluene at 110 ℃ for 18 h. The title compound (700 mg, 86%) was isolated as a solid by reverse phase chromatography (C18) using a gradient of 30% -100% MeCN and 10 mM ammonium formate in water. LCMS C ₃₈H₄₃N₅O₄ theory is 633.3 and found to be 635.9 [ m+h ] ⁺.

Step 2 Synthesis of N- [ (2R, 4R) -1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -2-methyl-4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester. To a solution of tert-butyl N- [ (2R, 4R) -1- [3- (2, 6-dibenzylox-3-pyridinyl) -1-methyl-indazol-6-yl ] -2-methyl-4-piperidinyl ] carbamate (800 mg,1.26 mmol) in DMF (13 mL) cooled to 0℃was added NaH (151 mg,3.8 mmol), and the reaction mixture was stirred at 0℃for 15min. Methyl iodide (0.24 mL,3.79 mmol) was added and the reaction mixture was warmed to room temperature and stirred 1 h. The reaction was diluted by dropwise addition of water (5 mL). Water (50 mL) and EtOAc (100 mL) were added and the layers separated. The organic layer was washed with water (5×20 mL), brine (50 mL), dried (MgSO ₄), filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (C18) using a gradient of 30% -100% MeCN and 10 mM ammonium formate in water to give the title compound as a solid (700 mg, 86%). LCMS C ₃₉H₄₅N₅O₄ theory 647.4 found 648.4 [ m+h ] ⁺.

Step 3 Synthesis of 3- (6- ((2R, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methylpiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was accomplished using general procedures 4, 5 and 6 with N- [ (2 r,4 r) -1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -2-methyl-4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester as starting material. The crude residue was purified by reverse phase chromatography (C18) using a gradient of 10% -70% MeCN and 10 mM ammonium formate in water to give the title compound as a solid (19 mg, 25%). LCMS C ₃₃H₃₆ClN₉O₃ theory 641.3, found 642.3[M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.88 (s, 1H), 8.60 (s, 1H), 8.02 (s, 1H), 7.70 (s, 1H), 7.60 (d, J = 8.7 Hz, 1H), 7.54 (s, 1H), 7.29 (s, 1H), 6.98 – 6.89 (m, 2H), 4.59 (br s, 1H), 4.33 (dd, J = 9.5, 4.9 Hz, 1H), 3.97 (s, 3H), 3.59 (s, 2H), 3.28 – 3.21 (m, 2H), 3.10 (s, 3H), 2.96 (s, 3H), 2.81 (br s, 1H), 2.68 – 2.59 (m, 2H), 2.38 – 2.31 (m, 1H), 2.22 – 2.13 (m, 1H), 1.95 – 1.86 (m, 1H), 1.76 (d, J = 10.6 Hz, 1H), 1.70 – 1.58 (m, 2H), 0.93 (d, J = 5.7 Hz, 3H).

EXAMPLE S5.Synthesis of 3- (6- (4- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) -2-methyl-6-oxopiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (5)

Step 1 synthesis of 6-methylpiperazin-2-one. To a stirred solution of 6-methylpiperazin-2-one (1.0 g,8.76 mmol) and triethylamine (1.83 mL,13.14 mmol) in CH ₂Cl₂ (20 mL) at 0℃under nitrogen was added Boc-anhydride (2.24 mL,9.64 mmol). The reaction mixture was slowly warmed to 25 ℃ and stirred 3 h. The reaction mixture was treated with 0.5N HCl (2.0 mL), water (50 mL) and extracted with DCM (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as an off-white solid (1.81 g,8.43 mmol,96% yield). The crude product was used in the next step without further purification.

Step 2 Synthesis of 4- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3-methyl-5-oxopiperazine-1-carboxylic acid tert-butyl ester. A stirred solution of 6-methylpiperazin-2-one (750 mg,3.49 mmol) and 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (1.50 g,3.00 mmol) in DMF (10 mL) was degassed with nitrogen 5 min. Copper (I) iodide (571 mg,3.00 mmol), K ₂CO₃ (621 mg,4.50 mmol) and N ₁,N₂ -dimethylethane-1, 2-diamine (264 mg,3.00 mmol) were then added and degassed 10 min. The reaction mixture was heated to 100 ℃ and stirred 16 h. The reaction mixture was cooled to room temperature and treated with water (30 mL) and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude compound. The crude product was purified by flash column chromatography on silica with 80% -100% ethyl acetate/petroleum ether to give the title compound as an off-white solid (650 mg,0.779 mmol,26% yield). MS (ESI) M/z 634.8 [ M+H ] ⁺.

Step 3 Synthesis of 3- (6- (4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methyl-6-oxopiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the synthesis of the title compound was started using general procedures 4,5 and 6 starting with 4- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3-methyl-5-oxopiperazine-1-carboxylic acid tert-butyl ester. LCMS C ₃₁H₃₀ClN₉O₄ theory 627.2, found 628.2[M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.92 (s, 1H), 8.83 (s, 1H), 8.10 (s, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.59 (s, 1H), 7.55-7.53 (m, 2H), 7.01 - 6.96 (m, 2H), 4.58 - 4.50 (m, 1H), 4.39 (dd, J = 5.1 Hz and 10.2 Hz, 1H), 4.24 - 4.16 (m, 2H), 4.25 - 4.07 (m, 1H), 3.98 (s, 3H), 3.90 - 3.83 (m, 1H), 3.55 (s, 2H), 3.12 (s, 3H), 2.73 - 2.66 (m, 2H), 2.66 - 2.61 (m, 1H), 2.43 - 2.36 (m, 1H), 1.07 (d, J = 6.3 Hz, 3H).

The examples in table 2 below were prepared according to the general procedure outlined in the table starting with their respective commercial starting materials and intermediates found in this document.

EXAMPLE S52.3 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (52)

Step 1 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. 3- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] piperidine-2, 6-dione (40.3 mg,0.120 mmol), 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (28.8 mg,0.100 mmol), N-diisopropylethylamine (0.03 mL,0.2000 mmol) and DMSO (0.1969 mL) were added to a 1-dram vial equipped with a stirring bar and heated to 80 ℃ overnight. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as an off-white solid (3.0 mg,0.0049 mmol,4.9% yield). LCMS C ₃₁H₃₂ClN₉O₃ theory was 613.2, found to be 614.2 [M+H]⁺;¹H NMR (DMSO-d₆, 400 MHz) δ 10.7-10.8 (m, 1H), 8.5-8.6 (m, 1H), 7.95 (s, 1H), 7.45 (s, 2H), 7.2-7.3 (m, 1H), 6.8-6.9 (m, 1H), 6.4-6.5 (m, 3H), 5.7-5.7 (m, 1H), 4.2-4.4 (m, 2H), 4.0-4.1 (m, 1H), 3.75 (s, 3H), 3.5-3.6 (m, 1H), 3.4-3.5 (m, 2H), 3.03 (s, 5H), 2.5-2.6 (m, 1H), 2.0-2.2 (m, 3H), 1.9-2.0 (m, 2H), 1.2-1.3 (m, 2H).

EXAMPLE S53 Synthesis of relative- (R) -3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (single isomer, unidentified stereochemistry) (53)

Step 1-Synthesis of relative- (R) -3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S52 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=11.0 min. LCMS C ₃₁H₃₂ClN₉O₃ theory 614.2, found 614.2 [M+H]⁺;¹H NMR (DMSO-d6, 400 MHz) δ 10.7-10.8 (m, 1H), 8.5-8.6 (m, 1H), 7.95 (s, 1H), 7.45 (s, 2H), 7.2-7.3 (m, 1H), 6.8-6.9 (m, 1H), 6.4-6.5 (m, 3H), 5.7-5.7 (m, 1H), 4.2-4.4 (m, 2H), 4.0-4.1 (m, 1H), 3.75 (s, 3H), 3.5-3.6 (m, 1H), 3.4-3.5 (m, 2H), 3.03 (s, 5H), 2.5-2.6 (m, 1H), 2.0-2.2 (m, 3H), 1.9-2.0 (m, 2H), 1.2-1.3 (m, 2H).

EXAMPLE S54 Synthesis of relative- (S) -3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (single isomer, unidentified stereochemistry) (54)

Step 1-Synthesis of relative- (S) -3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S52 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm, 5. Mu. Column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=18.9 min. LCMS C ₃₁H₃₂ClN₉O₃ theory 614.2, found 614.2 [M+H]⁺;¹H NMR (DMSO-d6, 400 MHz) δ 10.7-10.8 (m, 1H), 8.5-8.6 (m, 1H), 7.95 (s, 1H), 7.45 (s, 2H), 7.2-7.3 (m, 1H), 6.8-6.9 (m, 1H), 6.4-6.5 (m, 3H), 5.7-5.7 (m, 1H), 4.2-4.4 (m, 2H), 4.0-4.1 (m, 1H), 3.75 (s, 3H), 3.5-3.6 (m, 1H), 3.4-3.5 (m, 2H), 3.03 (s, 5H), 2.5-2.6 (m, 1H), 2.0-2.2 (m, 3H), 1.9-2.0 (m, 2H), 1.2-1.3 (m, 2H).

EXAMPLE S55.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-hydroxypiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione (55)

Step 1 Synthesis of 6-chloro-3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridine. To a solution of 6-chloro-3-iodo-1H-pyrazolo [3,4-b ] pyridine (1 g,3.58 mmol) in DMF (20 mL) at 0oC sodium hydride (0.150 g,3.76 mmol) was added and the reaction mixture stirred for 30 min. Sodium hydride (0.150 g,3.76 mmol) was then added dropwise. The reaction mixture was allowed to warm to room temperature and stirred for an additional hour. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite, and concentrated. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound as a white solid (0.75 g,2.56 mmol,71% yield ）;MS (ESI) m/z 293.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.59 (s, 1 H), 7.92 (s, 1 H), 4.06 (d, J=0.98 Hz, 3 H).

Step 2 Synthesis of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-chloro-1-methyl-1H-pyrazolo [3,4-b ] pyridine. A solution of 6-chloro-3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridine (500 mg,1.704 mmol), (2, 6-bis (benzyloxy) pyridin-3-yl) boronic acid (628 mg,1.874 mmol) and sodium carbonate (379 mg,3.58 mmol) in 1, 4-dioxane (6 mL) and water (2 mL) was stirred with tetrakis (triphenylphosphine) palladium (0) (197 mg,0.170 mmol) at 100 ℃. After this time, the reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite, and concentrated. The crude product was purified by column chromatography on silica gel (0-50% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound as a white solid (520 mg,1.14 mmol,67% yield ）;MS (ESI) m/z 456.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.25 (d, J=8.44 Hz, 1 H), 8.02 (d, J=8.07 Hz, 1 H), 7.45 - 7.51 (m, 2 H), 7.28 - 7.44 (m, 8 H), 7.11 (d, J=8.44 Hz, 1 H), 6.62 (d, J=8.19 Hz, 1 H), 5.49 (s, 2 H), 5.44 (s, 2 H), 4.04 (s, 3 H).

Step 3 Synthesis of tert-butyl (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-hydroxypiperidine-1-carboxylate. To a solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-chloro-1-methyl-1H-pyrazolo [3,4-b ] pyridine (500 mg,1.094 mmol), ruPhos Pd G3 (92 mg,0.109 mmol) and tert-butyl (3 r,4 r) -4-amino-3-hydroxypiperidine-1-carboxylate (237 mg,1.094 mmol) in 1, 4-dioxane (5 mL) was added cesium carbonate (713 mg,2.189 mmol). The reaction mixture was degassed with argon 1 min and then stirred 15 at 100 ℃ h. LCMS indicated completion of the reaction. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite, and concentrated. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give crude product. The crude product was further purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% TFA in water (0.1% TFA), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (120 mg,0.188 mmol,17% yield ）;MS (ESI) m/z 636.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.95 (d, J=8.07 Hz, 1 H), 7.65 (d, J=8.93 Hz, 1 H), 7.43 - 7.50 (m, 2 H), 7.28 - 7.43 (m, 8 H), 6.97 (d, J=7.09 Hz, 1 H), 6.56 (d, J=8.07 Hz, 1 H), 6.24 (d, J=8.93 Hz, 1 H), 5.46 (s, 2 H), 5.39 - 5.44 (m, 2 H), 5.18 (d, J=4.65 Hz, 1 H), 3.71 - 3.98 (m, 6 H), 3.40 (tt, J=9.00, 4.63 Hz, 1 H), 2.94 (br d, J=1.47 Hz, 1 H), 2.69 - 2.85 (m, 1 H), 1.99 - 2.15 (m, 1 H), 1.41 (s, 9 H), 1.24 - 1.34 (m, 1 H).

Step 4 Synthesis of 3- (6- (((3R, 4R) -3-hydroxypiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione. A solution of tert-butyl (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-hydroxypiperidine-1-carboxylate (0.120 g,0.188 mmol) in ethanol (10 mL) was stirred under hydrogen (1 atm) at 50℃with 10% w/w (0.020 g,0.019 mmol) palladium on carbon for 15H. The reaction mixture was filtered through celite, the filtrate was concentrated and then the residue was stirred with 1 mL TFA in DCM (1 mL) at room temperature for 30 min. Volatiles were removed under reduced pressure to give the desired product as TFA salt (67 mg,0.187 mmol,99% yield), which was used without further purification.

Step 5 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-hydroxypiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione. To a solution of 3- (6- (((3 r,4 r) -3-hydroxypiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione (68 mg,0.190 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (55.5 mg,0.190 mmol) in DMSO (3 mL) was added DIPEA (0.10 mL,0.57 mmol). The reaction mixture was stirred at 80 ℃ for 16 hours. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (3.7 mg,5.86 μmol,3.09% yield ）;MS (ESI) m/z 630.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.83 (s, 1 H), 8.68 (s, 1 H), 8.02 (s, 1 H), 7.65 (d, J=8.68 Hz, 1 H), 7.54 (s, 1 H), 7.50 (dd, J=8.44, 2.20 Hz, 1 H), 6.99 (br d, J=7.21 Hz, 1 H), 6.94 (d, J=8.44 Hz, 1 H), 6.37 (d, J=8.80 Hz, 1 H), 5.16 (dd, J=5.07, 1.77 Hz, 1 H), 4.39 - 4.53 (m, 1 H), 4.24 - 4.38 (m, 1 H), 4.14 (dd, J=9.35, 5.07 Hz, 1 H), 3.89 - 4.02 (m, 1 H), 3.77 (s, 3 H), 3.54 (s, 2 H), 3.44 (tt, J=9.25, 4.88 Hz, 1 H), 3.29 (s, 1 H), 3.11 (s, 3 H), 3.05 (br t, J=11.19 Hz, 1 H), 2.88 (dd, J=12.78, 9.60 Hz, 1 H), 2.55 - 2.64 (m, 2 H), 2.21 - 2.31 (m, 1 H), 2.06 - 2.20 (m, 2 H), 1.26 - 1.42 (m, 1 H).

EXAMPLE S56.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((3, 3-difluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (56)

Step 1, synthesis of 3, 3-difluoro-5-nitroindoline-2-ketone. To a suspension of 5-nitroindoline-2, 3-dione (2.2 g,11.5 mmol) in dichloromethane (50 mL) at-78 ℃ was added dropwise diethylaminosulfur trifluoride (4.06 g,25.2 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 2 days. LCMS indicated completion of the reaction. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried (anhydrous magnesium sulfate), filtered through celite and concentrated. The crude product was purified by silica gel column chromatography (0-75% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound 3, 3-difluoro-5-nitroindolin-2-one (1.52 g,7.10 mmol,62.0% yield) as a yellow solid ）;MS (ESI) m/z 215.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 11.89 (br s, 1 H), 8.56 - 8.60 (m, 1 H), 8.43 (d, J=8.78 Hz, 1 H), 7.20 (d, J=8.68 Hz, 1 H).

Step 2, synthesis of 3, 3-difluoro-1-methyl-5-nitroindoline-2-one. To a solution of 3, 3-difluoro-5-nitroindolin-2-one (0.53 g,2.475 mmol) in DMF (1 mL) was added DBU (0.485 mL,3.22 mmol) followed by methyl iodide (0.185 mL,2.97 mmol). The reaction mixture was stirred at room temperature for 15 hours. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 50 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried (anhydrous magnesium sulfate), filtered through celite and concentrated. The crude product was purified by column chromatography on silica gel (0-80% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound as a yellow solid (0.339 g,1.485 mmol,60% yield) ）;MS (ESI) m/z 229.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 8.61 (d, J=1.96 Hz, 1 H), 8.54 (d, J=8.91 Hz, 1 H), 7.46 (d, J=8.80 Hz, 1 H), 3.25 (s, 3 H).

Step 3 synthesis of 5-amino-3, 3-difluoro-1-methylindolin-2-one. A suspension of 3, 3-difluoro-1-methyl-5-nitroindolin-2-one (360 mg,1.578 mmol) in ethanol (15 mL) was stirred at room temperature under hydrogen (45 psi) for 15 h. The reaction mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give the title compound (300 mg,1.514 mmol,96% yield) as a yellow solid ）;MS (ESI) m/z 199.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 6.90 (d, J=8.44 Hz, 1 H), 6.87 (d, J=1.96 Hz, 1 H), 6.75 (d, J=7.91 Hz, 1 H), 5.22 (s, 2 H), 3.09 (s, 3 H).

Step 4 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((3, 3-difluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was completed using general procedures 1 and 6 using 5-amino-3, 3-difluoro-1-methylindolin-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride as starting materials. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a yellow solid (8 mg,0.012 mmol,23.46% yield ）;MS (ESI) m/z 664.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6) δ ppm 10.81 (s, 1 H), 8.94 (s, 1 H), 8.03 - 8.14 (m, 2 H), 7.77 (br d, J=8.56 Hz, 1 H), 7.31 (d, J=8.80 Hz, 1 H), 7.21 (d, J=8.44 Hz, 1 H), 6.51 (br d, J=8.80 Hz, 1 H), 6.44 (s, 1 H), 5.70 (br d, J=8.80 Hz, 1 H), 4.47 (br s, 2 H), 4.17 (dd, J=8.62, 5.07 Hz, 1 H), 3.81 (s, 3 H), 3.17 (s, 3 H), 3.04 (br t, J=12.10 Hz, 1 H), 2.65 - 2.76 (m, 1 H), 2.56 - 2.63 (m, 2 H), 2.19 - 2.31 (m, 1 H), 2.11 - 2.19 (m, 1 H), 2.06 (br d, J=10.64 Hz, 1 H), 1.46 - 1.66 (m, 1 H), 1.07 - 1.28 (m, 2 H), 0.96 (d, J=6.36 Hz, 3 H).

EXAMPLE S57.Synthesis of 3- [6- [ [ (3S, 4R) -1- [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -3- (hydroxymethyl) -4-piperidinyl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione (57)

Step 1 Synthesis of tert-butyl O5-ethyl 4- [ [ (1R) -1-phenylethyl ] amino ] -3, 6-dihydro-2H-pyridine-1, 5-dicarboxylic acid. To a solution of O1-tert-butyl O3-ethyl 4-oxopiperidine-1, 3-dicarboxylic acid ester (22 g,81 mmol) and (1R) -1-phenylethanamine (12.5 mL,97.2 mmol) in toluene (400 mL) was added PTSA (1.39 g,8.1 mmol) and the mixture was heated to reflux using a Dean-Stark trap (Dean-STARK TRAP) for 18 h. The mixture was cooled to room temperature and washed with saturated aqueous NaHCO ₃ (2 x 200 mL) and brine (2 x 200 mL). The combined organic fractions were dried over MgSO ₄, filtered and concentrated. The residue was filtered over a pad of silica, rinsed with DCM (2×100 mL) and concentrated to give the title compound as an oil (30.2 g, 99%) which was used in the next step without further purification .MS (ESI) [M+H]⁺ 375.2.¹H NMR (400 MHz, CDCl₃) δ 9.25 (d, J = 7.5 Hz, 1H), 7.36 – 7.28 (m, 2H), 7.26 – 7.18 (m, 3H), 4.66 – 4.54 (m, 1H), 4.24 – 4.14 (m, 2H), 4.07 (br s, 2H), 3.48 – 3.35 (m, 1H), 3.35 – 3.24 (m, 1H), 2.39 (dd, J = 13.0, 6.2 Hz, 1H), 2.09 – 2.00 (m, 1H), 1.50 (d, J = 6.8 Hz, 3H), 1.43 (s, 9H), 1.29 (t, J = 7.0 Hz, 3H).

Step 2 Synthesis of tert-ButylO 3-ethyl (3S, 4R) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1, 3-dicarboxylic acid ester and tert-ButylO 3-ethyl (3R, 4S) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1, 3-dicarboxylic acid ester sodium triacetoxyborohydride (34 g,160 mmol) was added in portions over 2H to a solution of tert-ButylO 5-ethyl 4- [ [ (1R) -1-phenylethyl ] amino ] -3, 6-dihydro-2H-pyridine-1, 5-dicarboxylic acid ester (15 g,40.1 mmol) in MeCN (200 mL) and AcOH (100 mL) cooled to 0℃and the reaction mixture was stirred at 0℃for 2H. The mixture was then cooled to-10 ℃ and slowly treated with 1M aqueous NaOH (100 mL), 4M aqueous NaOH (100 mL), 6M aqueous NaOH (100 mL), followed by 50% aqueous NaOH (50 mL). The mixture was warmed to room temperature and the layers were separated. The aqueous layer was extracted with DCM (3X 80 mL). The combined organic layers were dried over MgSO ₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 0-50% EtOAc in hexanes to give a 4:1 mixture of the title compound (14.8 g, 98%) as an oil. A portion of the 4:1 mixture of the title compound (7 g) was isolated by SFC to give tert-butyl O3-ethyl (3S, 4R) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1, 3-dicarboxylic acid ester (3.7 g) and the title compound tert-butyl O3-ethyl (3R, 4S) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1, 3-dicarboxylic acid ester (1.1 g) as an oil.

Step 3 Synthesis of tert-butyl (3S, 4R) -3- (hydroxymethyl) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylate. To a solution of tert-butyl O3-ethyl (3 s, 4R) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1, 3-dicarboxylic acid ester (500 mg,1.33 mmol) in THF (3 mL) was added LiBH ₄ (1.33 mL,2.66 mmol) and the mixture was heated to reflux for 2h and then cooled to room temperature. Ice-water (10 mL) was added and the mixture was concentrated under reduced pressure. The residue was extracted with EtOAc (3X 30 mL). The combined organic fractions were dried over Na ₂SO₄, filtered and concentrated to give the title compound as an oil (404 mg, 91%) which was used in the next step without further purification. MS (ESI) [ M+H ] ⁺ 335.3.

Step 4 Synthesis of tert-butyl (3S, 4R) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylate. To a solution of tert-butyl (3 s, 4R) -3- (hydroxymethyl) -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylate (404 mg,1.21 mmol) in DCM (5 mL) was added tert-butylchlorodimethylsilane (218 mg,1.45 mmol) and imidazole (123 mg,1.81 mmol) in sequence and the reaction mixture was stirred at room temperature for 3h. Water (15 mL) and Et ₂ O (20 mL) were added and the layers separated. The aqueous layer was washed with Et ₂ O (3 x 10 mL) and the combined organic fractions were dried over Na ₂SO₄, filtered and concentrated to give the title compound as an oil (540 mg, 99%) which was used in the next step without further purification. Note that SFC analysis confirmed the presence of only one diastereomer .MS (ESI) [M+H]⁺ 449.4.¹H NMR (400 MHz, DMSO-d₆) δ 7.35 – 7.26 (m, 4H), 7.23 – 7.16 (m, 1H), 3.86 – 3.68 (m, 3H), 3.68 – 3.54 (m, 1H), 3.48 (t, J = 9.6 Hz, 1H), 2.89 – 2.68 (m, 1H), 2.81 (dd, J = 13.1, 3.2 Hz, 1H), 2.59 – 2.52 (m, 1H), 1.82 – 1.72 (m, 1H), 1.36 (s, 9H), 1.34 – 1.27 (m, 1H), 1.26 – 1.18 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H), 0.90 (s, 9H), 0.07 (d, J = 3.1 Hz, 6H).

Step 5 Synthesis of (3S, 4R) -4-amino-3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] piperidine-1-carboxylic acid tert-butyl ester. A mixture of (3S, 4R) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylic acid tert-butyl ester (530 mg,1.18 mmol), ammonium formate (596 mg,9.45 mmol) and Pd/C (126 mg,0.12 mmol) in EtOH (15 mL) was heated to 65℃for 2 h. The mixture was filtered through celite and washed with MeOH (3×15 mL). The filtrate was concentrated under reduced pressure to give the title compound (320 mg, 79%) as a solid, which was used in the next step without further purification .MS (ESI) [M+H]⁺ 345.3.¹H NMR (400 MHz, DMSO-d₆) δ 3.61 (dd, J = 10.2, 4.9 Hz, 1H), 3.49 – 3.41 (m, 2H), 3.39 – 3.24 (m, 3H), 3.23 – 3.08 (m, 1H), 3.06 – 2.98 (m, 1H), 1.71 – 1.55 (m, 2H), 1.55 – 1.44 (m, 1H), 1.40 – 1.33 (m, 1H), 1.38 (s, 9H), 0.89 – 0.84 (m, 9H), 0.06 – 0.01 (m, 6H).

Step 6 (3S, 4R) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylic acid tert-butyl ester synthesis. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (471 mg,0.94 mmol), (3S, 4R) -4-amino-3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] piperidine-1-carboxylic acid tert-butyl ester (270 mg,0.78 mmol), cs ₂CO₃ (638 mg,1.96 mmol) and RuPhos Pd G (98.3 mg,0.12 mmol) in 1, 4-dioxane (7 mL) was heated to 90℃for 16 h and then cooled to room temperature. The residue was purified by column chromatography on silica gel using a gradient of 0-50% EtOAC in hexane to give the title compound as a solid (458 mg, 67%). Note that the reaction was repeated and the crude residue was combined prior to purification .MS (ESI) [M+H]⁺ 764.4.¹H NMR (400 MHz, DMSO-d₆) δ 7.87 (d, J = 8.1 Hz, 1H), 7.50 – 7.44 (m, 2H), 7.42 – 7.26 (m, 9H), 6.55 (d, J = 7.9 Hz, 1H), 6.52 (dd, J = 9.0, 1.9 Hz, 1H), 6.45 (d, J = 1.3 Hz, 1H), 5.78 (d, J = 8.6 Hz, 1H), 5.44 (s, 2H), 5.41 (s, 2H), 3.88 (s, 3H), 3.85 – 3.77 (m, 1H), 3.67 – 3.61 (m, 1H), 3.60 – 3.41 (m, 4H), 3.39 – 3.32 (m, 1H), 2.10 – 2.00 (m, 1H), 1.67 – 1.53 (m, 2H), 1.41 (s, 9H), 0.82 (s, 9H), –0.05 (d, J = 15.0 Hz, 6H).

Step 7 synthesis of (3S, 4R) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylic acid tert-butyl ester. A mixture of (3S, 4R) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylic acid tert-butyl ester (458 mg,0.60 mmol) and Peltier catalyst (210 mg,0.15 mmol) in EtOH (10 mL) and THF (10 mL) was subjected to hydrogenation (1 atm) 2h at 50 ℃. The mixture was filtered through celite and washed with MeOH (3×15 mL). The filtrate was concentrated under reduced pressure to give the title compound (350 mg, quantitative) as a solid, which was used in the next step without further purification. MS (ESI) [ M+H ] ⁺ 586.4.

Step 8 Synthesis of 3- [6- [ [ (3S, 4R) -3- (hydroxymethyl) -4-piperidinyl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione dihydrochloride. To a solution of tert-butyl (3 s,4 r) -3- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate (350 mg,0.60 mmol) in DCM (2.5 mL) was added 4N HCl (1.5 ml,6.0 mmol) in 1, 4-dioxane and the mixture stirred at room temperature for 2 h. The resulting precipitate was collected by filtration, washed with Et ₂ O (3 x 5 mL) and dried under vacuum to give the title compound (250 mg, 77%) as a solid, which was used in the next step without further purification .MS (ESI) [M+H]⁺ 372.2.¹H NMR (500 MHz, DMSO-d₆) δ 10.82 (s, 1H), 8.89 – 8.68 (m, 2H), 7.35 (d, J = 8.8 Hz, 1H), 6.66 (dd, J = 8.8, 1.7 Hz, 1H), 6.51 (s, 1H), 4.18 (dd, J = 9.0, 5.1 Hz, 1H), 3.99 – 3.92 (m, 1H), 3.82 (s, 3H), 3.51 – 3.41 (m, 2H), 3.27 – 3.15 (m, 2H), 3.15 – 3.04 (m, 2H), 2.67 – 2.54 (m, 2H), 2.32 – 2.21 (m, 2H), 2.18 – 2.10 (m, 1H), 1.98 – 1.89 (m, 1H), 1.87 – 1.78 (m, 1H).

Step 9 Synthesis of 3- [6- [ [ (3S, 4R) -1- [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -3- (hydroxymethyl) -4-piperidinyl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione. To a mixture of 5- [ (5-chloro-2-fluoro-pyrimidin-4-yl) amino ] -1-methyl-indolin-2-one (35 mg,0.12 mmol) and DIPEA (0.10 mL,0.60 mmol) in DMF (2 mL) was added 3- [6- [ [ (3 s,4 r) -3- (hydroxymethyl) -4-piperidinyl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione dihydrochloride (77.8 mg,0.14 mmol) at room temperature, and the reaction mixture was heated to 80 ℃ for 2 h. Volatiles were removed under reduced pressure and the residue was purified by preparative HPLC (BEH column, C18) using a gradient of 31% -41% MeCN and 10 mM ammonium formate in water to give the title compound (22.1 mg, 28%) as a solid. LCMS C ₃₂H₃₄ClN₉O₄ theory 643.2, found 644.4 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.81 (s, 1H), 8.61 (s, 1H), 8.00 (s, 1H), 7.75 – 7.57 (m, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 6.65 (d, J = 8.9 Hz, 1H), 6.48 (s, 1H), 5.85 (d, J = 8.5 Hz, 1H), 4.46 (s, 1H), 4.18 (dd, J = 8.4, 5.1 Hz, 1H), 3.96 – 3.85 (m, 2H), 3.81 (s, 3H), 3.80 – 3.75 (m, 1H), 3.75 – 3.63 (m, 2H), 3.52 (s, 2H), 3.51 – 3.46 (m, 1H), 3.09 (s, 3H), 2.66 – 2.55 (m, 2H), 2.31 – 2.21 (m, 1H), 2.20 – 2.11 (m, 1H), 2.11 – 2.02 (m, 1H), 1.79 – 1.69 (m, 1H), 1.67 – 1.56 (m, 1H).

EXAMPLE S58.Synthesis of 3- (6- (((3R, 4S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (hydroxymethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (58)

Step 1 Synthesis of 3- (6- (((3R, 4S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (hydroxymethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized in analogy to example S57, starting with the other diastereoisomers isolated from example S57 step 2. The crude reaction mixture was purified by preparative HPLC (BEH column, C18) using a gradient of 44% -54% MeCN and 10mM ammonium formate in water to give the title compound (5.5 mg, 14%) as a solid. LCMS C ₃₂H₃₄ClN₉O₄ theory 643.2, found 644.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.00 (s, 1H), 7.66 (br s, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.33 (d, J = 8.8 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 6.65 (dd, J = 8.8, 1.7 Hz, 1H), 6.48 (s, 1H), 5.85 (d, J = 8.5 Hz, 1H), 4.18 (dd, J = 8.8, 5.1 Hz, 1H), 3.92 – 3.84 (m, 2H), 3.81 (s, 3H), 3.80 – 3.74 (m, 1H), 3.52 (s, 2H), 3.51 – 3.46 (m, 2H), 3.39 – 3.29 (m, 2H), 3.09 (s, 3H), 2.63 – 2.57 (m, 2H), 2.30 – 2.19 (m, 1H), 2.19 – 2.11 (m, 1H), 2.11 – 2.01 (m, 1H), 1.78 – 1.69 (m, 1H), 1.67 – 1.52 (m, 1H).

EXAMPLE S59.Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) piperidin-1-yl) -1-methyl-1H-pyrazolo [4,3-b ] pyridin-3-yl) piperidine-2, 6-dione (59)

Step 1 Synthesis of 6-bromo-3-iodo-1H-pyrazolo [4,3-b ] pyridine. To a solution of 6-bromo-1H-pyrazolo [4,3-b ] pyridine (3.75 g,18.9 mmol) in MeCN (90 mL) was added NIS (5.11 g,22.7 mmol). The reaction mixture was heated to 85 ℃ for 18 h and then cooled to room temperature. The volatiles were evaporated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-15% EtOAc in hexanes to give the title compound (6.05 g, 98%) as a solid. MS (ESI) [ M+H ] ⁺ 324.0.0.

Step 2 Synthesis of 6-bromo-3-iodo-1-methyl-pyrazolo [4,3-b ] pyridine. The title compound was synthesized in analogy to general procedure 3, using 6-bromo-3-iodo-1H-pyrazolo [4,3-b ] pyridine as starting material.

Step 3 Synthesis of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-b ] pyridine. A mixture of 6-bromo-3-iodo-1-methyl-pyrazolo [4,3-b ] pyridine (900 mg,2.66 mmol), (2, 6-dibenzyloxy-3-pyridinyl) boronic acid (5.36 g,7.99 mmol, intermediate provided by Celgene), pd (PPh ₃)₂Cl₂ (374 mg,0.53 mmol) and Na ₂CO₃ (2.66 mL,5.33 mmol) in 1, 4-dioxane (18 mL) in 2M aqueous solution was heated to 80 ℃ for 16H and then cooled to room temperature.

Step 4 Synthesis of tert-butyl N- [1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-pyrazolo [4,3-b ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamate. The title compound was synthesized according to general procedure 2 using 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-b ] pyridine and N-methyl-N- (4-piperidinyl) carbamic acid tert-butyl ester as starting materials in toluene.

Step 5 Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) piperidin-1-yl) -1-methyl-1H-pyrazolo [4,3-b ] pyridin-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was accomplished using general procedures 4, 5 and 6 using N- [1- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-b ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester as starting material. The crude reaction mixture was purified by preparative HPLC (BEH column, C18) using a gradient of 44% -54% MeCN and 10 mM ammonium formate in water to give the title compound (5.4 mg, 12%) as a solid. LCMS C ₃₁H₃₃ClN₁₀O₃ theory 628.2, found 629.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.89 – 10.82 (m, 1H), 8.59 (s, 1H), 8.41 (d, J = 2.3 Hz, 1H), 8.03 (s, 1H), 7.65 (s, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 2.3 Hz, 1H), 6.95 (d, J = 8.6 Hz, 1H), 4.28 (dd, J = 9.8, 5.2 Hz, 1H), 3.97 – 3.91 (m, 5H), 3.55 (s, 2H), 3.28 (s, 2H), 3.08 (s, 3H), 2.92 (s, 3H), 2.87 – 2.72 (m, 2H), 2.72 – 2.64 (m, 1H), 2.62 – 2.55 (m, 1H), 2.22 – 2.10 (m, 1H), 1.97 – 1.84 (m, 2H), 1.76 – 1.66 (m, 2H).

EXAMPLE S60.Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) piperidin-1-yl) -1-methyl-1H-pyrazolo [4,3-c ] pyridin-3-yl) piperidine-2, 6-dione (60)

Step 1 6-bromo-3-iodo-1H-pyrazolo [4,3-c ] pyridine to a solution of 6-bromo-1H-pyrazolo [4,3-c ] pyridine (3.5 g,17.7 mmol) in MeCN (177 mL) was added NIS (7.95 g,35.4 mmol). The reaction mixture was heated to 85 ℃ for 18 h and then cooled to room temperature. Water (50 mL) was added and the resulting precipitate was collected by filtration, washed with water (50 mL), DCM (50 mL), and dried under vacuum to give the title compound (6.21 g, quantitative) as a solid ）.MS (ESI) [M+H]⁺ 325.8.¹H NMR (400 MHz, DMSO-d₆) δ 8.44 (s, 1H), 7.57 (s, 1H).

Step 2 6-bromo-3-iodo-1-methyl-pyrazolo [4,3-c ] pyridine. To a mixture of 6-bromo-3-iodo-1H-pyrazolo [4,3-c ] pyridine (100 mg,0.31 mmol) in DMF (2 mL) was added NaH (60% dispersion in mineral oil, 31 mg,0.77 mmol) at 0 ℃ and the reaction mixture was stirred 15 min at 0 ℃. A solution of methyl iodide (50 μl,0.77 mmol) in DMF (1 mL) was added and the reaction warmed to room temperature and stirred 2 h. Water (20 mL) and DCM (20 mL) were added and the layers separated. The aqueous layer was extracted with DCM (2X 20 mL). The combined organic fractions were washed with brine (40 mL), dried over MgSO ₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 10% -35% EtOAc in hexanes to give the title compound as a solid （46 mg,44%）.¹H NMR (400 MHz, DMSO-d₆) δ 8.58 (d, J = 1.0 Hz, 1H), 8.09 (d, J = 1.0 Hz, 1H), 4.05 (s, 3H).

Step 3 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-c ] pyridine. A mixture of 6-bromo-3-iodo-1-methyl-pyrazolo [4,3-c ] pyridine (100 mg,0.30 mmol), (2, 6-dibenzyloxy-3-pyridinyl) boronic acid (595 mg,0.89 mmol), pd (PPh ₃)₂Cl₂ (41.5 mg,60 μmol) and Na ₂CO₃ (300 μL,0.59 mmol) in 1, 4-dioxane (1.5 mL) in 2M aqueous solution was heated to 80 ℃ for 16 h and then cooled to room temperature, water (40 mL) and EtOAc (40 mL) were added and the layers were separated （99.0 mg,67%）.¹H NMR (400 MHz, DMSO-d₆) δ 8.87 (d, J = 1.0 Hz, 1H), 8.09 – 7.96 (m, 2H), 7.49 – 7.44 (m, 2H), 7.43 – 7.39 (m, 2H), 7.38 – 7.33 (m, 3H), 7.33 – 7.24 (m, 3H), 6.63 (d, J = 8.2 Hz, 1H), 5.52 (s, 2H), 5.44 (s, 2H), 4.06 (s, 3H).

Step 4N- [1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-pyrazolo [4,3-c ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-c ] pyridine (1.44G, 2.87 mmol), tert-butyl N-methyl-N- (4-piperidinyl) carbamate (677 mg,3.16 mmol), ruPhos-Pd-G3 (480 mg,0.57 mmol) and NaOtBu (552 mg,5.74 mmol) in 1, 4-dioxane (58 mL) was heated to 90℃for 16 h and then cooled to room temperature. The volatiles were evaporated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-40% EtOAc in hexanes to give the title compound as a solid （1.08 g,59%）.¹H NMR (400 MHz, DMSO-d₆) δ 8.71 (d, J = 1.0 Hz, 1H), 7.97 (d, J = 8.1 Hz, 1H), 7.51 – 7.43 (m, 2H), 7.43 – 7.33 (m, 5H), 7.33 – 7.23 (m, 3H), 6.72 (d, J = 1.1 Hz, 1H), 6.58 (d, J = 8.1 Hz, 1H), 5.51 (s, 2H), 5.40 (s, 2H), 4.47 (d, J = 12.6 Hz, 2H), 3.93 (s, 3H), 2.82 (t, J = 12.5 Hz, 2H), 2.72 – 2.66 (m, 1H), 2.64 (s, 3H), 1.80 – 1.52 (m, 4H), 1.40 (s, 9H).

Step 5N- [1- [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-pyrazolo [4,3-c ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester. A mixture of N- [1- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-pyrazolo [4,3-C ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester (1.59 g,1.88 mmol) and Pd (OH) ₂/C (200 mg,0.38 mmol) in THF (12.5 mL) and EtOH (12.5 mL) was subjected to hydrogenation (1 atm) 2h at room temperature. The mixture was filtered through celite and washed with DMF (60 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound (927 mg, quantitative) as a solid. MS (ESI) [ M+H ] ⁺ 457.4.

Step 6 3- [ 1-methyl-6- [4- (methylamino) -1-piperidinyl ] pyrazolo [4,3-c ] pyridin-3-yl ] piperidine-2, 6-dione hydrochloride. To a solution of N- [1- [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-pyrazolo [4,3-c ] pyridin-6-yl ] -4-piperidinyl ] -N-methyl-carbamic acid tert-butyl ester (927 mg,2.03 mmol) in DCM (40.6 mL) was added 4N HCl (5.08 mL,20.3 mmol) in 1, 4-dioxane and the reaction mixture was stirred at room temperature 16 h. The reaction was diluted with Et ₂ O (20 mL), filtered and washed with Et ₂ O (2 x 10 mL) to give the title compound (688 mg, 86%) as a solid which was used in the next step without further purification .MS (ESI) [M+H]⁺ 357.3.¹H NMR (400 MHz, DMSO-d₆) δ 10.99 (s, 1H), 9.06 (s, 2H), 8.91 (s, 1H), 7.17 (s, 1H), 4.51 (dd, J = 10.9, 4.9 Hz, 1H), 4.33 (d, J = 13.3 Hz, 2H), 3.97 (s, 3H), 3.36 – 3.17 (m, 1H), 3.07 (t, J = 12.6 Hz, 2H), 2.77 – 2.60 (m, 2H), 2.56 (t, J = 5.4 Hz, 3H), 2.48 – 2.38 (m, 1H), 2.25 – 2.15 (m, 1H), 2.14 (d, J = 13.4 Hz, 2H), 1.75 – 1.57 (m, 2H).

Step 7 3- [6- [4- [ [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -methyl-amino ] -1-piperidinyl ] -1-methyl-pyrazolo [4,3-c ] pyridin-3-yl ] piperidine-2, 6-dione to a solution of 3- [ 1-methyl-6- [4- (methylamino) -1-piperidinyl ] pyrazolo [4,3-c ] pyridin-3-yl ] piperidine-2, 6-dione hydrochloride (50 mg,0.13 mmol) in DMSO (1.6 mL- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (34 mg,0.12 mmol) and DIPEA (0.2 mL,1.16 mmol) were added sequentially. The reaction mixture was heated to 100 ℃ for 18 h and then cooled to room temperature. The crude reaction mixture was purified by preparative HPLC (BEH column, C18) using a gradient of 44% -54% MeCN and 10mM ammonium formate in water to give the title compound (18.6 mg, 25%) as a solid. LCMS C ₃₁H₃₃ClN₁₀O₃ theory 628.2, found 629.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.90 (s, 1H), 8.64 (s, 1H), 8.59 (s, 1H), 8.11 – 7.84 (m, 1H), 7.64 (s, 1H), 7.58 (d, J = 8.4 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 6.74 (s, 1H), 4.53 (d, J = 12.9 Hz, 2H), 4.35 – 4.25 (m, 1H), 3.98 – 3.82 (m, 3H), 3.54 (s, 1H), 3.30 – 3.26 (m, 3H), 3.09 (s, 2H), 2.91 –2.73 (m, 4H), 2.73 – 2.54 (m, 3H), 2.41 – 2.29 (m, 1H), 2.23 – 2.13 (m, 1H), 1.82 – 1.59 (m, 4H).

EXAMPLE S61.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-6-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (61)

Step1, synthesis of 1-methyl-6-nitroindoline-2, 3-dione. To a stirred solution of 6-nitroindoline-2, 3-dione (1.0 g,5.20 mmol) in DMF (20 mL) under nitrogen at 0 ℃ was added sodium hydride (0.312 g,7.81 mmol) and stirred 10 min. Methyl iodide (0.651 mL,10.41 mmol) was added to the reaction mixture and warmed slowly to 25 ℃. The reaction mixture was stirred for 1h, treated with saturated NH ₄ Cl solution (100 mL) and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound, which was purified by flash column chromatography on silica gel with 40% -50% ethyl acetate/petroleum ether to give the title compound as an off-white solid (320 mg,1.531 mmol,29% yield).

Step 2 synthesis of 1-methyl-6-nitroindolin-2-one. To a stirred solution of 1-methyl-6-nitroindoline-2, 3-dione (200 mg,0.957 mmol) in n-butanol (5.0 mL) was added hydrazine hydrate (144 mg,2.87 mmol) at 25 ℃. The reaction was heated to 80 ℃ and stirred 4 h. The reaction mixture was then cooled to 25 ℃ and triethylamine (0.533 mL,3.83 mmol) was added and stirred at 80 ℃ 14 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude compound, which was purified by flash column chromatography on silica gel with 35% -45% ethyl acetate/petroleum ether to give the title compound as an off-white solid (40 mg,0.171 mmol,18% yield). LCMS found 193.2 [ M+H ] ⁺, rt 1.686 min

Step 3, synthesis of 6-amino-1-methylindolin-2-one. To a stirred solution of 1-methyl-6-nitroindolin-2-one (40 mg,0.171 mmol) in ethanol (2.5 mL) and THF (2.5 mL) at 25 ℃ under nitrogen was added 10% Pd/C (20 mg). The reaction mixture was stirred under a hydrogen atmosphere for 6 h. The reaction mixture was filtered through celite and the celite pad was washed with ethanol (2×20 mL). The filtrate was concentrated under reduced pressure to give the title compound (35 mg, crude), which was used in the next step without further purification .¹H NMR (400 MHz, CDCl₃): δ 6.86 (d, J = 8.4 Hz, 1H), 6.20-6.18 (m, 2H), 5.11 (brs, 2H), 3.32 (s, 2H), 3.02 (s, 3H).

Step 4 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-6-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was accomplished using general procedures 1 and 6 using 6-amino-1-methylindolin-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. The crude material was purified by preparative HPLC to give the title compound as an off-white solid (18 mg,0.028 mmol,17% yield). LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.84 (s, 1H), 8.99 (s, 1H), 8.12 (s, 1H), 7.42 (s, 1H), 7.32 (d, J = 8.80 Hz, 1H), 7.26 (d, J = 9.20 Hz, 1H), 7.21 (d, J = 8.00 Hz, 1H), 6.52 (dd, J = 8.80, 1.60 Hz, 1H), 6.45 (s, 1H), 5.79 (brs, 1H), 4.55-4.47 (m, 2H), 4.19-4.16 (m, 1H), 3.81 (s, 3H), 3.12-3.06 (m, 4H), 2.75-2.70 (m, 1H), 2.66-2.58 (m, 3H), 2.28-2.19 (m, 1H), 2.16-2.08 (m, 2H), 1.62-1.53 (m, 1H), 1.25-1.18 (m, 1H), 0.97 (d, J = 6.4 Hz, 3H).

EXAMPLE S62.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione (62)

Step 1.6-chloro-3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridine sodium hydride (0.301 g,7.51 mmol) was added to a 0 ℃ solution of 6-chloro-3-iodo-1H-pyrazolo [3,4-b ] pyridine (2 g,7.16 mmol) in DMF (20 mL), the reaction mixture was stirred for 30 min and then sodium hydride (0.301 g,7.51 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred 1 h. The reaction mixture was treated with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography (0-100% ethyl acetate in hexanes). The desired fractions were concentrated under reduced pressure to give the title compound 6-chloro-3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridine (1.7 g,5.79 mmol,81% yield) as a white solid ）;MS (ESI) m/z 293.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.99 (d, J=8.31 Hz, 1 H), 7.32 (d, J=8.31 Hz, 1 H), 4.03 (s, 3 H).

(3R, 4R) -4- ((3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester to a solution of 6-chloro-3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridine (500 mg,1.704 mmol) and N-ethyl-N-isopropyl-propan-2-amine (0.607 mL,3.41 mmol) in DMSO (1 mL) was added (3R, 4R) -4-amino-3-methylpiperidine-1-carboxylic acid tert-butyl ester (365 mg,1.704 mmol). The reaction mixture was stirred at 120 ℃ for 48 h. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (250 mg,0.530 mmol,31.1% yield ）.MS (ESI) m/z 472.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.33 (dd, J=8.68, 1.71 Hz, 1 H), 7.18 (br d, J=7.83 Hz, 1 H), 6.33 - 6.46 (m, 1 H), 3.86 - 4.05 (m, 2 H), 3.81 (d, J=1.83 Hz, 4 H), 2.79 - 2.99 (m, 1 H), 2.56 - 2.64 (m, 1 H), 1.98 (br d, J=12.35 Hz, 1 H), 1.47 - 1.63 (m, 1 H), 1.41 (d, J=1.71 Hz, 9 H), 1.17 - 1.32 (m, 1 H), 0.88 (br d, J=4.89 Hz, 3 H).

(3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester. A solution of (3R, 4R) -4- ((3-iodo-1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester (250 mg,0.530 mmol), 2, 6-bis (benzyloxy) -3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridine (243 mg,0.583 mmol) and sodium bicarbonate (98 mg,1.167 mmol) in a mixed solvent of 1, 4-dioxane (10 mL) and water (2 mL) was stirred with tetrakis (triphenylphosphine) palladium (0) (61.3 mg,0.053 mmol) at 80 ℃. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fraction was concentrated under reduced pressure to give the crude product which was further purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (110 mg,0.173 mmol,32.7% yield ）.MS (ESI) m/z 372.0 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.94 (d, J=8.07 Hz, 1 H), 7.64 (d, J=8.80 Hz, 1 H), 7.43 - 7.51 (m, 2 H), 7.36 - 7.42 (m, 4 H), 7.26 - 7.36 (m, 4 H), 6.92 (d, J=8.19 Hz, 1 H), 6.55 (d, J=8.19 Hz, 1 H), 6.19 (d, J=8.93 Hz, 1 H), 5.46 (s, 2 H), 5.41 (s, 2 H), 3.86 - 3.99 (m, 2 H), 3.81 - 3.85 (m, 3 H), 3.69 - 3.81 (m, 1 H), 2.75 - 3.07 (m, 1 H), 1.91 - 2.07 (m, 1 H), 1.45 - 1.64 (m, 1 H), 1.41 (s, 9 H), 1.24 - 1.32 (m, 2 H), 0.88 (d, J=6.60 Hz, 3 H).

Step 4.3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione. A solution of tert-butyl (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylate (0.105 g,0.165 mmol) in ethanol (10 mL) was stirred under hydrogen (1 atm) at 50℃with 10% w/w (0.018 g,0.017 mmol) palladium on carbon for 15H. The reaction mixture was filtered through celite, the filtrate was concentrated and then the residue was stirred with 1 mL TFA in DCM (1 mL) at room temperature for 30 min. TFA and solvent were removed under reduced pressure to give the desired product 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione (60 mg,0.168 mmol, quantitative yield) as TFA salt, which was used without further purification.

Step 5.3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione the synthesis of the title compound was completed using general procedure 6 using 3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-pyrazolo [3,4-b ] pyridin-3-yl) piperidine-2, 6-dione hydrochloride (40 mg,0.102 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (29.8 mg,0.102 mmol). The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid （13 mg,0.02 mmol,19.4%）.MS (ESI) m/z 628.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.83 (s, 1 H), 8.66 (s, 1 H), 8.02 (s, 1 H), 7.63 (d, J=8.80 Hz, 1 H), 7.57 (s, 1 H), 7.52 (dd, J=8.44, 2.08 Hz, 1 H), 6.87 - 6.98 (m, 2 H), 6.31 (d, J=8.80 Hz, 1 H), 4.48 (br s, 2 H), 4.13 (dd, J=9.35, 5.07 Hz, 1 H), 3.82 - 3.95 (m, 1 H), 3.77 (s, 3 H), 3.53 (s, 2 H), 3.06 - 3.15 (m, 3 H), 2.92 - 3.03 (m, 1 H), 2.64 - 2.73 (m, 1 H), 2.52 - 2.64 (m, 2 H), 2.21 - 2.31 (m, 1 H), 2.09 - 2.19 (m, 1 H), 2.02 (br d, J=9.29 Hz, 1 H), 1.54 - 1.67 (m, 1 H), 1.21 - 1.37 (m, 1 H), 0.93 (d, J=6.48 Hz, 3 H).

EXAMPLE S63.3 Synthesis of 3- (6- ((1- (5-chloro-4- ((2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (63)

Step 1 Synthesis of 3- (6- ((1- (5-chloro-4- ((2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized in analogy to example S62 described above, using 3- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] piperidine-2, 6-dione and 5-aminoindolin-2-one as starting materials. LCMS C ₃₀H₃₀ClN₉O₃ theory 599.2, found 600.0 [M+H]⁺;¹H NMR (DMSO-d₆, 400 MHz) δ 10.8-10.9 (m, 1H), 10.3-10.4 (m, 1H), 8.0-8.1 (m, 1H), 7.4-7.5 (m, 1H), 7.3-7.4 (m, 2H), 6.8-6.8 (m, 1H), 6.4-6.6 (m, 2H), 4.2-4.4 (m, 2H), 4.1-4.2 (m, 1H), 3.9-4.0 (m, 2H), 3.84 (s, 3H), 3.6-3.7 (m, 1H), 3.49 (s, 2H), 3.1-3.2 (m, 2H), 2.6-2.6 (m, 2H), 2.2-2.3 (m, 1H), 2.1-2.2 (m, 1H), 2.0-2.1 (m, 2H), 1.3-1.4 (m, 2H).

EXAMPLE S64.3 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (64)

Step 1 Synthesis of 3- (6- ((3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride. A mixture of 3- (6-amino-1-methyl-indazol-3-yl) piperidine-2, 6-dione (500 mg,1.94 mmol), 3-difluoro-4-oxopiperidine-1-carboxylic acid tert-butyl ester hydrate (500.9 mg,2.13 mmol) and decaborane (14) (108.54 mg,0.9700 mmol) was stirred in DMSO (5 mL) and acetic acid (0.22 mL,3.87 mmol) at ambient temperature. After 2 hours, the reaction was completed according to LCMS. The reaction was diluted with methanol (1 mL), which resulted in bubbling. After 10 min, the bubbling subsided and the solution was diluted with ethyl acetate, washed with water and then brine, dried (sodium sulfate), filtered and concentrated. The resulting solid was triturated with diethyl ether. The resulting solid was then purified using silica gel chromatography using a gradient of 4% methanol and 0-50% ethyl acetate in hexane. The resulting intermediate 4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] -3, 3-difluoro-piperidine-1-carboxylic acid tert-butyl ester (230 mg,0.4817 mmol,24.8% yield) was isolated and treated with 4N HCL in 1, 4-dioxane of 2 mL and stirred 3 h. After this time, the reaction mixture was concentrated to give the title compound (270 mg,0.6524 mmol,33.7% yield) as a pale yellow powder.

Step 2 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione 3- {6- [ (3, 3-difluoropiperidin-4-yl) amino ] -1-methyl-1H-indazol-3-yl } piperidine-2, 6-dione hydrochloride (278 mg,0.650 mmol), 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (172 mg,0.590 mmol), N-diisopropylethylamine (0.2 mL,1.17 mmol) and DMSO (1.17 mL) were added to a 1-dram vial equipped with a stirring rod and heated to 80℃for 15 hours. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound (12 mg,0.0184 mmol,3.14% yield ）.¹H NMR (DMSO-d₆, 400 MHz) δ 10.8-10.9 (m, 1H), 8.8-8.9 (m, 1H), 8.2-8.2 (m, 1H), 8.0-8.1 (m, 1H), 7.4-7.5 (m, 2H), 7.3-7.4 (m, 1H), 6.9-7.0 (m, 1H), 6.6-6.7 (m, 2H), 5.9-6.1 (m, 1H), 4.6-4.8 (m, 1H), 4.4-4.5 (m, 1H), 4.2-4.3 (m, 2H), 3.8-3.9 (m, 3H), 3.4-3.6 (m, 3H), 3.2-3.3 (m, 1H), 3.1-3.1 (m, 3H), 2.6-2.7 (m, 2H), 2.5-2.6 (m, 4H), 2.2-2.3 (m, 1H), 2.1-2.2 (m, 1H), 1.9-2.0 (m, 1H), 1.5-1.7 (m, 1H).

EXAMPLE S65.Synthesis of 1- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (65)

Step 1 Synthesis of tert-butyl 4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] -3, 3-difluoro-piperidine-1-carboxylate. To a solution of 1- (6-amino-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione (900 mg,3.47 mmol), 3-difluoro-4-oxo-piperidine-1-carboxylic acid tert-butyl ester (980 mg,4.17 mmol) in acetic acid (5 mL) and DMSO (25 mL) was added decaborane (14) (195 mg,1.74 mmol) and the mixture stirred at room temperature for 6 h. Volatiles were removed and the residue was purified by reverse phase chromatography (C18) with a gradient of 20% -100% MeCN and 10 mM ammonium formate in water to give the title compound as a solid (450 mg, 27%). MS (ESI) [ M-H ] ^- 477.3.

Step 2 Synthesis of 1- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedures 5 and 6 using 4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] -3, 3-difluoro-piperidine-1-carboxylic acid tert-butyl ester as starting material. Theoretical LCMS C ₃₀H₂₉ClF₂N₁₀O₃ of 650.2, found 651.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.47 (s, 1H), 8.82 (s, 1H), 8.06 (s, 1H), 7.51 (d, J = 2.1 Hz, 1H), 7.47 (dd, J = 8.4, 2.2 Hz, 1H), 7.31 (d, J = 8.8 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.65 (dd, J = 8.9, 1.9 Hz, 1H), 6.61 (d, J = 1.8 Hz, 1H), 6.05 (d, J = 9.2 Hz, 1H), 4.77 – 4.63 (m, 1H), 4.49 – 4.36 (m, 1H), 4.33 – 4.17 (m, 1H), 3.87 (t, J = 6.7 Hz, 2H), 3.81 (s, 3H), 3.55 – 3.51 (m, 2H), 3.58 – 3.44 (m, 1H), 3.28 – 3.18 (m, 1H), 3.11 (s, 3H), 2.72 (t, J = 6.7 Hz, 2H), 2.03 – 1.95 (m, 1H), 1.70 – 1.57 (m, 1H).

EXAMPLE S66.3 Synthesis of 3- (6- (((R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (66)

Step 1 Synthesis of tert-butyl 3, 3-difluoro-4- (((R) -1-phenylethyl) amino) piperidine-1-carboxylate. A mixture of 3, 3-difluoro-4-oxo-piperidine-1-carboxylic acid tert-butyl ester (3.53 g,15.0 mmol) and (1R) -1-phenylethanamine (2.18 g,18.0 mmol) in toluene (35 mL) was heated to reflux using a dean-Stark trap for 18: 18 h. The mixture was cooled to room temperature and volatiles were removed under reduced pressure. DCM (35 mL) was added followed by sodium triacetoxyborohydride (7.15 g,33.7 mmol) in portions over 1:1 h. The mixture was heated to 50 ℃ for 18 h. Another batch of sodium triacetoxyborohydride (3.56 g,16.8 mmol) was added and the reaction mixture was heated to 50 ℃ for a further 6 h. The mixture was cooled to room temperature and volatiles were removed under reduced pressure. A solution of Na ₂CO₃ (50 mL) in 2M was added and the mixture was stirred at room temperature for 1 h, followed by EtOAc (50 mL). The phases were separated and the aqueous phase was extracted with EtOAc (3×50 mL). The combined organic fractions were washed with brine (50 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using 0-3% MeOH in DCM to give the title compound as an oil (4.21 g, 82%). ¹ H NMR (500 MHz, methanol -d₄) δ 7.29 – 7.35 (m, 4H), 7.20 – 7.25 (m, 1H), 5.48 (s, 1H), 4.08 (q, J = 6.7 Hz, 1H), 4.04 (br s, 1 H), 3.73 – 3.79 (m, 1H), 3.17 (br s, 1H), 2.91 (br s, 1H), 2.70 – 2.80 (m, 1H), 1.65 – 1.72 (m, 1H), 1.45 – 1.50 (m, 1H), 1.44 (s, 9H), 1.31 (d, J = 6.62 Hz, 3H).)

Step 2 Synthesis of (4R) -3, 3-difluoro-4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylic acid tert-butyl ester 4-methylbenzenesulfonate. To a solution of tert-butyl 3, 3-difluoro-4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylate (4.21 g,12.4 mmol) in EtOH (50 mL) was added 4-methylbenzenesulfonic acid hydrate (2.35 g,12.4 mmol) at room temperature and the mixture was heated to 60 ℃ for 1 h. Volatiles were removed under reduced pressure to give a diastereomeric mixture as a solid. The crude mixture of intermediates (6.07 g) was dissolved in a 10:1 mixture of acetone: etOH (68.5 mL) and the mixture was heated to reflux. The flask was removed from the oil bath and left to stand overnight. The precipitated solid was filtered, rinsed with cold acetone (2 x 1 mL) and dried to give the title compound which was first harvested, which was taken to the subsequent process.

Step 3 Synthesis of (4R) -3, 3-difluoro-4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylic acid tert-butyl ester. The compound (4R) -3, 3-difluoro-4- [ [ (1R) -1-phenylethyl ] amino ] piperidine-1-carboxylic acid tert-butyl ester 4-methylbenzenesulfonate (1.99 g,3.88 mmol) was partitioned between EtOAc (50 mL) and saturated aqueous NaHCO3 (50 mL). The organic layer was separated and the aqueous phase extracted with EtOAc (3×40 mL). The combined organic layers were dried (MgSO 4), filtered and concentrated under reduced pressure to give the title compound as an oil （1.30 g,98%）.MS (ESI) [M+H]⁺ 341.3.¹H NMR (400 MHz, DMSO-d6) δ 7.36 – 7.27 (m, 4H), 7.26 – 7.15 (m, 1H), 4.01 – 3.95 (m, 1H), 3.94 – 3.80 (m, 1H), 3.66 – 3.54 (m, 1H), 3.29 – 3.20 (m, 1H), 3.04 – 2.91 (m, 1H), 2.71 – 2.64 (m, 1H), 2.37 (t, J = 7.2 Hz, 1H), 1.64 – 1.58 (m, 1H), 1.38 (s, 9H), 1.22 (d, J = 6.6 Hz, 3H).

Step 4 Synthesis of (4R) -4-amino-3, 3-difluoro-piperidine-1-carboxylic acid tert-butyl ester. A mixture of tert-butyl (4R) -3, 3-difluoro-4- [ [ (1 SR) -1-phenylethyl ] amino ] piperidine-1-carboxylate (1.29 g,3.79 mmol) and 10% Pd/C (202 mg,0.19 mmol) in EtOH (41 mL) was shaken in a Parr flask at room temperature under hydrogen atmosphere (50 psi) for 18 h. The mixture was filtered over celite and washed with MeOH (3×50 mL). The filtrate was concentrated under reduced pressure to give the title compound (845 mg, 94%) as an oil, which was used in the next step without further purification. MS (ESI) [ M-butene ] +181.0. [ alpha ] D+0.19° (c=0.142, etOH). ¹ H NMR (400 MHz, methanol -d4) δ 4.22 – 4.12 (m, 1H), 3.96 (dt, J = 13.7, 4.0 Hz, 1H), 3.25 – 3.15 (m, 1H), 3.14 – 3.00 (m, 2H), 1.93 – 1.82 (m, 1H), 1.59 – 1.48 (m, 1H), 1.46 (s, 9H).)

Step 5 (4R) -Synthesis of tert-butyl 4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] -3, 3-difluoro-piperidine-1-carboxylate. To a degassed mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (200 mg,0.40 mmol), (4R) -4-amino-3, 3-difluoro-piperidine-1-carboxylic acid tert-butyl ester (123 mg,0.520 mmol) and tBuONa (57.7 mg,0.60 mmol) in THF (2 mL) was added tbux pos-Pd-G3 (33.9 mg,40 μmol) at room temperature. The reaction vessel was sealed and the reaction mixture was heated to 80 ℃ for 18 h and cooled to room temperature. DCM (10 mL) was added and the mixture was filtered over celite and rinsed with DCM (3×10 mL). The volatiles were concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-60% EtOAc in hexanes to give the title compound as a solid （200 mg,76%）.MS (ESI) [M+H]⁺ 657.6.¹H NMR (400 MHz, DMSO-d₆) δ 7.86 (d, J = 8.1 Hz, 1H), 7.50 – 7.43 (m, 2H), 7.42 – 7.23 (m, 9H), 6.59 (s, 1H), 6.58 – 6.52 (m, 2H), 6.00 (d, J = 9.1 Hz, 1H), 5.44 (s, 2H), 5.41 (s, 2H), 4.25 – 4.08 (m, 2H), 3.97 – 3.92 (m, 1H), 3.89 (s, 3H), 3.50 – 3.36 (m, 1H), 3.19 – 3.00 (m, 1H), 1.97 – 1.87 (m, 1H), 1.65 – 1.52 (m, 1H), 1.42 (s, 9H).

Step 6 Synthesis of 3- (6- (((R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 4, 5 and 6 using (4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] -3, 3-difluoro-piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₁H₃₀ClF₂N₉O₃ theory 649.2, found 650.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 8.82 (s, 1H), 8.41 (s, 1H), 8.06 (s, 1H), 7.51 (s, 1H), 7.47 (dd, J = 8.4, 2.1 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.66 (dd, J = 8.8, 1.8 Hz, 1H), 6.62 (s, 1H), 6.00 (d, J = 8.9 Hz, 1H), 4.75 – 4.64 (m, 1H), 4.41 (d, J = 11.6 Hz, 1H), 4.26 – 4.13 (m, 2H), 3.82 (s, 3H), 3.60 – 3.42 (m, 3H), 3.23 (t, J = 11.7 Hz, 1H), 3.11 (s, 3H), 2.65 – 2.55 (m, 2H), 2.30 – 2.22 (m, 1H), 2.20 – 2.11 (m, 1H), 2.04 – 1.94 (m, 1H), 1.70 – 1.52 (m, 1H).

EXAMPLE S67.Synthesis of 3- (6- (((S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (67)

Step 1 Synthesis of 3- (6- (((S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3, 3-difluoropiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the title compound was synthesized in analogy to example S66 using (1S) -1-phenylethylamine in step 1. LCMS C ₃₁H₃₀ClF₂N₉O₃ theory 649.2, found 650.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.81 (s, 1H), 8.82 (s, 1H), 8.06 (s, 1H), 7.51 (s, 1H), 7.48 (d, J = 8.3 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.66 (d, J = 8.4 Hz, 1H), 6.62 (s, 1H), 6.00 (d, J = 9.0 Hz, 1H), 4.78 – 4.63 (m, 1H), 4.47 – 4.33 (m, 1H), 4.30 – 4.18 (m, 2H), 3.82 (s, 3H), 3.60 – 3.45 (m, 3H), 3.26 – 3.19 (m, 1H), 3.11 (s, 3H), 2.69 – 2.57 (m, 2H), 2.31 – 2.23 (m, 1H), 2.15 (dd, J= 12.8, 5.9 Hz, 1H), 2.05 – 1.92 (m, 1H), 1.70 – 1.56 (m, 1H).

EXAMPLE S68.Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) amino) -3, 3-difluoropiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (68)

Step 1 Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) amino) -3, 3-difluoropiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 2, 4, 5 and 6 using tert-butyl (3, 3-difluoropiperidin-4-yl) carbamate as starting material. LCMS C ₃₁H₃₀ClF₂N₉O₃ theory 649, found 650 [M+H]⁺;¹H NMR (DMSO-d₆, 400 MHz) δ 10.8-10.9 (m, 1H), 8.5-8.7 (m, 1H), 7.9-8.1 (m, 1H), 7.6-7.8 (m, 1H), 7.4-7.6 (m, 2H), 7.2-7.4 (m, 1H), 6.9-7.0 (m, 3H), 4.2-4.4 (m, 1H), 4.0-4.2 (m, 1H), 3.92 (s, 4H), 3.56 (s, 2H), 3.3-3.3 (m, 1H), 3.1-3.2 (m, 3H), 3.0-3.1 (m, 1H), 2.9-2.9 (m, 1H), 2.7-2.7 (m, 1H), 2.6-2.7 (m, 1H), 2.2-2.3 (m, 1H), 2.1-2.2 (m, 1H), 1.9-2.0 (m, 2H).

Example S69.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-ethyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (69)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-ethyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1-ethylindol-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₃H₃₆ClN₉O₃ theory 641.3, found 642.4 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ ppm 10.82 (s, 1H), 9.03 (br s, 1H), 8.09 (s, 1H), 7.55 (s, 1H), 7.50 (dd, J = 8.50, 1.90 Hz, 1H), 7.31 (d, J = 8.68 Hz, 1H), 7.01 (d, J = 8.44 Hz, 1H), 6.51 (dd, J = 8.68, 1.59 Hz, 1H), 6.45 (s, 1H), 5.54 - 6.00 (m, 1H), 4.39 (br d, J = 7.70 Hz, 2H), 4.17 (dd, J = 8.68, 5.14 Hz, 1H), 3.81 (s, 3H), 3.68 (q, J = 7.09 Hz, 2H), 3.54 (s, 2H), 3.25 - 3.37 (m, 1H), 3.08 (br t, J = 12.10 Hz, 1H), 2.75 (br t, J = 12.41 Hz, 1H), 2.54 - 2.64 (m, 2H), 2.20 - 2.31 (m, 1H), 2.04 - 2.19 (m, 2H), 1.52 - 1.69 (m, 1H), 1.16 - 1.27 (m, 1H), 1.13 (t, J = 7.09 Hz, 3 H), 0.97 (d, J = 6.48 Hz, 3H).

EXAMPLE S70.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (70)

Step 1 Synthesis of (3R, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester (intermediate 24). A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (1.92 g,3.84 mmol), (3R, 4R) -4-amino-3-methyl-piperidine-1-carboxylic acid tert-butyl ester (685 mg,3.20 mmol), ruPhos Pd G (668.6 mg,0.80 mmol), and Cs ₂CO₃ (1.25 g,3.84 mmol) in 1, 4-dioxane (15 mL) was heated to 90℃for 20 h. The mixture was cooled to room temperature, filtered through celite and washed with EtOAc (4×30 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-40% EtOAc in hexanes to give the title compound as a solid （1.37 g,68%）.MS (ESI) [M+H]⁺ 634.1.¹H NMR (400 MHz, DMSO-d₆) δ 7.86 (d, J = 8.1 Hz, 1H), 7.50 – 7.46 (m, 1H), 7.46 – 7.25 (m, 10H), 6.54 (d, J = 8.1 Hz, 1H), 6.43 (d, J = 1.8 Hz, 1H), 6.41 (d, J = 2.2 Hz, 1H), 5.70 (d, J = 8.9 Hz, 1H), 5.44 (s, 2H), 5.40 (s, 2H), 3.97 – 3.89 (m, 2H), 3.88 (s, 3H), 3.24 – 3.14 (m, 1H), 2.99 – 2.87 (m, 1H), 2.05 – 2.01 (m, 1H), 2.01 – 1.98 (m, 1H), 1.57 – 1.47 (m, 1H), 1.41 (s, 9H), 1.17 – 1.08 (m, 1H), 0.93 (d, J = 6.5 Hz, 3H).

Step 2 Synthesis of tert-butyl (3R, 4R) -4- [ [3- (2, 6-dioxo-3-piperidyl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylate. A mixture of (3R, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester (1.40 g,2.21 mmol) and Pd (OH) ₂/C (1.18 g,1.10 mmol) in MeOH (25 mL) and THF (75 mL) was hydrogenated at 50℃at H ₂ (1 atm) to 9H. The mixture was filtered through celite and washed with MeOH (2 x 50 mL) and THF (2 x 100 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-5% MeOH in DCM to give the title compound as a solid （695 mg,69%）.MS (ESI) [M+H]⁺ 456.3.¹H NMR (400 MHz, DMSO-d₆) δ 10.81 (s, 1H), 7.31 (d, J = 8.8 Hz, 1H), 6.51 (dd, J = 8.8, 1.7 Hz, 1H), 6.40 (s, 1H), 5.72 (d, J = 8.9 Hz, 1H), 4.17 (dd, J = 8.7, 5.2 Hz, 1H), 3.98 – 3.88 (m, 2H), 3.80 (s, 3H), 3.26 – 3.14 (m, 1H), 3.00 – 2.86 (m, 1H), 2.65 – 2.56 (m, 3H), 2.34 – 2.20 (m, 1H), 2.18 – 2.09 (m, 1H), 2.05 – 1.94 (m, 1H), 1.60 – 1.46 (m, 1H), 1.41 (s, 9H), 1.22 – 1.05 (m, 1H), 0.92 (d, J = 6.5 Hz, 3H).

Step 3 Synthesis of 3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride. To a solution of (3 r,4 r) -4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester (8 g,17.6 mmol) in ethyl acetate (35 mL) was added 4M HCl (35 mL) in ethyl acetate at 25 ℃. After addition, the reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was filtered to give the title compound as an off-white solid (8 g,100% yield ）.MS (ESI) [M+H]⁺356.2.400 MHz MeOD δ: 8.04 (s, 1H), 7.81 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 8.8 Hz, 1H), 4.59-4.55 (m, 1H), 4.11 (s, 3H), 3.75-7.71 (m, 1H), 3.49-3.46 (m,2 H), 3.20-3.15 (m, 1H), 3.03 (s, 2H), 2.93-2.89 (m, 1H), 2.83-2.81 (m, 2H), 2.52-2.48 (m, 1H), 2.40-2.35 (m, 1H), 2.30-2.26 (m, 1H), 2.20-2.15 (m, 1H), 2.02 (s, 1H), 1.80-1.75 (m, 1H), 1.24-1.22 (m, 3H).

Step 4 Synthesis of 3- [6- [ [ (3R, 4R) -1- [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -3-methyl-4-piperidinyl ] amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione. To a solution of 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride (50.0 mg,0.130 mmol) in DMSO (0.50 mL) was added sequentially 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (41.1 mg,0.140 mmol) and DIPEA (90 μl,0.510 mmol) and heated to 80 ℃ for 2H. The reaction mixture was cooled to room temperature and purified directly by preparative HPLC (BEH column, C18) using a gradient of 44% -54% MeCN and 10mM ammonium formate in water to give the title compound (13.5 mg, 16%) as a solid. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.81 (s, 1H), 8.65 (s, 1H), 8.02 (s, 1H), 7.57 (s, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.93 (d, J = 8.4 Hz, 1H), 6.50 (d, J = 8.8 Hz, 1H), 6.43 (s, 1H), 5.68 (d, J = 9.1 Hz, 1H), 4.53 – 4.39 (m, 2H), 4.17 (dd, J = 8.7, 5.2 Hz, 1H), 3.81 (s, 3H), 3.53 (s, 2H), 3.27 – 3.21 (m, 1H), 3.10 (s, 3H), 3.02 (t, J = 12.2 Hz, 1H), 2.76 – 2.65 (m, 1H), 2.64 – 2.56 (m, 2H), 2.32 – 2.19 (m, 1H), 2.18 – 2.09 (m, 1H), 2.10 – 2.02 (m, 1H), 1.65 – 1.51 (m, 1H), 1.21 – 1.11 (m, 1H), 0.97 (d, J = 6.5 Hz, 3H).

EXAMPLE S71 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (71)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S70 was then isolated using preparative HPLC (with REGIS WHELK-O1 RR or SS column, flow rate of 1 mL/min, isocratic elution in 100% ACN (no additives)) at 22 ℃. The title compound was eluted as the first peak, rt=10.01 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (DMSO-d₆, 400 MHz) δ 10.7-10.9 (m, 1H), 8.5-8.8 (m, 1H), 7.9-8.1 (m, 1H), 7.5-7.7 (m, 2H), 7.2-7.4 (m, 1H), 6.9-7.1 (m, 1H), 6.4-6.7 (m, 2H), 5.6-5.7 (m, 1H), 4.4-4.7 (m, 2H), 4.0-4.3 (m, 1H), 3.8-3.9 (m, 3H), 3.4-3.6 (m, 2H), 2.9-3.2 (m, 4H), 2.7-2.8 (m, 2H), 2.0-2.3 (m, 5H), 1.5-1.6 (m, 1H), 1.1-1.3 (m, 1H), 0.8-1.1 (m, 3H).

EXAMPLE S72 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (72)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S70 was then isolated using preparative HPLC (with REGIS WHELK-O1 RR or SS column, 1 mL/min flow rate, isocratic elution in 100% ACN (no additive) at 22 ℃. The title compound was eluted as the second peak with rt=14.87 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (DMSO-d6, 400 MHz) δ 10.7-10.9 (m, 1H), 8.6-8.8 (m, 1H), 7.9-8.1 (m, 1H), 7.5-7.7 (m, 2H), 7.2-7.4 (m, 1H), 6.9-7.0 (m, 1H), 6.3-6.6 (m, 2H), 5.6-5.9 (m, 1H), 4.4-4.6 (m, 2H), 4.1-4.4 (m, 1H), 3.8-3.9 (m, 3H), 3.5-3.6 (m, 2H), 2.9-3.2 (m, 4H), 2.7-2.8 (m, 2H), 2.0-2.3 (m, 5H), 1.5-1.7 (m, 1H), 1.1-1.3 (m, 1H), 0.9-1.0 (m, 3H)

EXAMPLE S73.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (73)

Step 1 Synthesis of (3R, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester. To an ice-cold solution of (3 r,4 r) -4- [ [3- (2, 6-dibenzylox-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester (1.55 g,2.45 mmol) and 60% NaH dispersion in mineral oil (245 mg,6.11 mmol) in DMF (20 mL) was added methyl iodide (305 μl,4.89 mmol) and the mixture was stirred at room temperature for 48 h. Another batch was prepared using the same procedure. Water (50 mL) was added to the combined mixture and the layers were separated. The aqueous layer was extracted with EtOAc (3X 75 mL). The combined organic fractions were washed with brine (20 mL), dried over MgSO ₄, filtered and concentrated. The residue was purified by reverse phase chromatography (C18) using a gradient of 20% -100% MeCN and 10 mM ammonium formate in water to give the title compound as a solid （700 mg,44%）.MS (ESI) [M+H]⁺ 649.34.¹H NMR (400 MHz, DMSO-d₆) δ 7.88 (d, J = 8.1 Hz, 1H), 7.47 (dd, J = 8.7, 7.1 Hz, 3H), 7.43 – 7.23 (m, 8H), 6.78 (dd, J = 9.4, 2.0 Hz, 1H), 6.61 (d, J = 1.9 Hz, 1H), 6.56 (d, J = 8.1 Hz, 1H), 5.44 (s, 2H), 5.40 (s, 2H), 4.05 – 3.99 (m, 2H), 3.94 (s, 3H), 3.65 – 3.59 (m, 1H), 2.92 – 2.85 (m, 1H), 2.75 (s, 3H), 2.60 – 2.54 (m, 1H), 1.72 – 1.77 (m, 1H), 1.60 – 1.54 (m, 2H), 1.42 (s, 9H), 0.75 (d, J = 6.5 Hz, 3H).

Step 2 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound is synthesized according to general procedures 4, 5 and 6 using (3 r,4 r) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₂H₃₄ClN₉O₃ theory 641.3, found 642.4 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.80 (s, 1H), 8.62 (s, 1H), 7.98 (s, 1H), 7.55 (s, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 9.1 Hz, 1H), 6.90 (d, J = 8.5 Hz, 1H), 6.85 (d, J = 7.4 Hz, 1H), 6.59 (s, 1H), 4.58 – 4.51 (m, 2H), 4.19 (dd, J = 8.9, 5.2 Hz, 1H), 3.83 (s, 3H), 3.74 – 3.65 (m, 1H), 3.49 (s, 2H), 3.07 (s, 3H), 2.97 – 2.90 (m, 1H), 2.69 (s, 3H), 2.63 – 2.57 (m, 3H), 2.30 – 2.23 (m, 1H), 2.16 – 2.09 (m, 1H), 1.86 – 1.76 (m, 1H), 1.60 – 1.53 (m, 2H), 0.77 (d, J = 6.5 Hz, 3H).

EXAMPLE S74.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxo-2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (74)

Step 1 Synthesis of 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methyl-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one. To a stirred solution of 5-amino-1-methyl-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one (1.2 g,1.18 mmol) in EtOH (30 mL) at-25 ℃ was added DIPEA (0.617 mL,3.53 mmol) and 5-chloro-2, 4-difluoropyrimidine (0.266 g,1.765 mmol). After 1 h the reaction was allowed to slowly warm to room temperature and then the reaction was stirred at 25 ℃ below 16: 16 h. After this time, the reaction mixture was evaporated. The crude residue was purified by silica gel chromatography using 50% -100% EtOAc in PE to give the title compound as a pale yellow solid (190 mg,0.252 mmol,21.44% yield). MS (ESI) M/z 294.0 [ M+H ] ⁺.

Step 2 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxo-2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. To a stirred solution of 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methyl-1, 3-dihydro-2H-pyrrolo [2,3-b ] pyridin-2-one (80 mg,0.104 mmol) and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione (49.1 mg,0.104 mmol) in DMSO (1 mL) was added DIPEA (0.054 mL,0.311 mmol) and the reaction mixture was stirred at 90 ℃ for 4H. The reaction mixture was purified by preparative HPLC to give the title compound as an off-white solid (30 mg,0.047 mmol,45.8% yield). LCMS C ₃₁H₃₃ClN₁₀O₃ theory 628.2, found 629.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6): δ 10.82 (s, 1H), 8.85 (s, 1H), 8.36 (d, J = 2.00 Hz, 1H), 8.06 (s, 1H), 7.86 (s, 1H), 7.31 (d, J = 8.80 Hz, 1H), 6.44-6.52 (m, 2H), 5.69 (d, J = 8.80 Hz, 1H), 4.44 (d, J = 9.20 Hz, 2H), 4.17 (q, J = 5.20 Hz, 1H), 3.82 (s, 3H), 3.61 (s, 2H), 3.13 (s, 3H), δ 3.02 (t, J = Hz, 1H), 2.53-2.68 (m, 3H), 2.33 (t, J = 1.60 Hz, 1H), 2.25 (t, J = 6.00 Hz, 1H), 2.06-2.17 (m, 2H), 1.58 (d, J = 4.80 Hz, 1H), 1.15-1.17 (m, 1H), 0.96 (d, J = 6.40 Hz, 3H).

EXAMPLE S75.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (75)

Step 1 Synthesis of 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1- (2-methoxyethyl) indolin-2-one. To a stirred solution of 5-amino-1- (2-methoxyethyl) indol-2-one (200 mg,0.97 mmol) in THF (5 mL) at-45 ℃ under nitrogen was added DIPEA (0.254 mL,1.46 mmol) and 5-chloro-2, 4-difluoropyrimidine (219 mg,1.46 mmol). The reaction mixture was slowly warmed to 25 ℃ and stirred 12 h. The reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography on silica gel with 30% ethyl acetate/petroleum ether to give the title compound as an off-white solid (80 mg,0.210 mmol,21% yield). MS: 337.0 [ M+H ] ⁺.

Step 2 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. To a stirred solution of 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione (117 mg,0.238 mmol) in DMSO (2.0 mL) was added 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1- (2-methoxyethyl) indolin-2-one (80 mg,0.238 mmol) and DIPEA (0.166 mL,0.95 mmol) at 25 ℃. The reaction mixture was heated to 80 ℃ and stirred 4 h. The crude product was purified by prep HPLC to give the title compound (24 mg,0.028 mmol,25% yield). LCMS C ₃₄H₃₈ClN₉O₄ theory 671.3, found 672.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6): δ 10.83 (s, 1H), 8.67 (s, 1H), 8.02 (s, 1H), 7.56 (s, 1H), 7.50 (d, J = 8.40 Hz, 1H), 7.31 (d, J = 8.80 Hz, 1H), 7.01 (d, J = 8.40 Hz, 1H), 6.51 (dd, J = 1.6 Hz and 8.8 Hz, 1H), 6.44 (s, 1H), 5.71 (d, j=9.20 Hz, 1H), 4.51-4.44 (m, 2H), 4.18 (dd, j=5.2 Hz and 8.4 Hz, 1H), 3.84-3.82 (m, 2H), 3.82 (s, 3H), 3.56-3.52 (m, 4H), 3.22 (s, 3H), 3.05-2.99 (m, 1H), 2.68-2.65 (m, 1H), 2.62-2.58 (m, 2H), 2.28-2.23 (m, 1H), 2.17-2.06 (m, 2H), 1.60-1.57 (m, 1H), 1.18-1.16 (m, 1H), 0.97 (d, J = 6.40 Hz, 3H).

EXAMPLE S76 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (76)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S75 was then isolated as a first elution peak using preparative HPLC (with REGIS WHELK-O1 SS,21 x 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% ACN (no additive) at 22 ℃. LCMS C ₃₄H₃₈ClN₉O₄ theory 671.3, found 672.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) = 10.81 (s, 1H), 8.65 (s, 1H), 8.02 (s, 1H), 7.56 (s, 1H), 7.49 (dd, J = 2.0, 8.5 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 6.50 (dd, J = 1.7, 8.8 Hz, 1H), 6.43 (d, J = 1.1 Hz, 1H), 5.69 (d, J = 8.9 Hz, 1H), 4.53 - 4.40 (m, 2H), 4.17 (dd, J = 5.2, 8.7 Hz, 1H), 3.86 - 3.79 (m, 5H), 3.58 - 3.48 (m, 4H), 3.21 (s, 3H), 3.06 - 2.96 (m, 1H), 2.73 - 2.56 (m, 3H), 2.35 - 2.01 (m, 4H), 1.63 - 1.50 (m, 1H), 1.23 - 1.10 (m, 1H), 0.96 (d, J = 6.6 Hz, 3H).

EXAMPLE S77 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (77)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-methoxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S75 was then isolated as a second elution peak using preparative HPLC (with Whelk-O1 SS,21 x 250 mm,5 u column, flow rate, 1 mL/min, isocratic elution in 100% ACN (no additive) at 22 ℃. LCMS C ₃₄H₃₈ClN₉O₄ theory 671.3, found 672.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) = 10.81 (s, 1H), 8.65 (s, 1H), 8.02 (s, 1H), 7.56 (s, 1H), 7.49 (dd, J = 2.1, 8.4 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 6.50 (dd, J = 1.7, 8.8 Hz, 1H), 6.43 (d, J = 1.2 Hz, 1H), 5.69 (d, J = 9.2 Hz, 1H), 4.52 - 4.41 (m, 2H), 4.17 (dd, J = 5.2, 8.7 Hz, 1H), 3.86 - 3.79 (m, 5H), 3.58 - 3.48 (m, 4H), 3.21 (s, 3H), 3.07 - 2.96 (m, 1H), 2.73 - 2.56 (m, 3H), 2.34 - 2.02 (m, 4H), 1.63 - 1.53 (m, 1H), 1.22 - 1.10 (m, 1H), 0.97 (d, J = 6.5 Hz, 3H).

EXAMPLE S78.3 Synthesis of 3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (78)

Step 1 Synthesis of (3R, 5R) -4- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3, 5-dimethylpiperazine-1-carboxylic acid tert-butyl ester. A stirred solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (2.335 g,4.67 mmol) and tert-butyl (3R, 5R) -3, 5-dimethylpiperazine-1-carboxylate (1.0 g,4.67 mmol) in toluene (25 ml) was degassed with nitrogen 15 min. Sodium tert-butoxide (0.673 g,7.00 mmol) and CPhos-Pd-G3 (0.188 g,0.233 mmol) were then added. The reaction mixture was heated to 100 ℃ and stirred 8 h. The reaction mixture was cooled to room temperature and treated with water (150 mL) and extracted with ethyl acetate (3×100 ml). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude compound. The crude product was purified by flash column chromatography on silica with 35% -40% ethyl acetate/petroleum ether to give the title compound as an off-white solid (950 mg,1.364 mmol,29% yield). MS (ESI) M/z 634.8 [ M+H ] ⁺.

Step 2 Synthesis of tert-butyl (3R, 5R) -4- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -3, 5-dimethylpiperazine-1-carboxylate. To a stirred solution of tert-butyl (3 r,5 r) -4- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3, 5-dimethylpiperazine-1-carboxylate (950 mg,1.364 mmol) in ethanol (20 mL) and THF (20 mL) under nitrogen was added 20% palladium on carbon hydroxide (192 mg). The reaction mixture was stirred at 25 ℃ under hydrogenation pressure 8 h. The reaction mixture was filtered through a pad of celite and the pad was washed with a 1:1 mixture of acetonitrile and methanol (3×25 mL). The filtrate was concentrated under reduced pressure to give the crude compound. The crude product was purified by flash column chromatography on silica with 5% MeOH/DCM to give the title compound as an off-white solid (330 mg,0.719 mmol,52.7% yield). MS (ESI) M/z 456.2 [ M+H ] ⁺.

Step 3 Synthesis of 3- (6- ((2R, 6R) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione TFA salt. To a stirred solution of tert-butyl (3 r,5 r) -4- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -3, 5-dimethylpiperazine-1-carboxylate (250 mg,0.450 mmol) in DCM (5 mL) at 0 ℃ was added TFA (0.34 mL). The reaction mixture was slowly warmed to 25 ℃ and stirred 4 h. The reaction mixture was concentrated under reduced pressure to give a crude compound. The crude compound was triturated with ether and dried under reduced pressure to give the title compound as an off-white solid (220 mg,0.410 mmol,91% yield). The crude product was used in the next step without further purification. MS (ESI) M/z 356.0 [ M+H ] ⁺.

Step 4 Synthesis of 3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. To a stirred solution of 3- (6- ((2 r,6 r) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione TFA salt (70 mg,0.137 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (40.2 mg,0.137 mmol) in DMSO (1 mL) at 25 ℃. The reaction mixture was heated to 80 ℃ and stirred for 6h. The reaction mixture was cooled to room temperature and purified by preparative HPLC to give the title compound as an off-white solid (26 mg,0.040 mmol,29% yield). LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6): δ 10.88 (s, 1H), 8.72 (s, 1H), 8.04 (s, 1H), 7.60 (s, 1H), 7.57-7.52 (m, 2H), 7.01 (s, 1H), 6.96 (d, J = 8.40 Hz, 1H), 6.87 (d, J = 8.80 Hz, 1H), 4.31-4.27 (m, 1H), 3.92 (s, 3H), 3.93-3.90 (m, 2H), 3.81-3.77 (m, 2H), 3.69-3.65 (m, 2H), 3.57 (s, 2H), 3.12 (s, 3H), 2.67-2.64 (m, 2H), 2.33-2.30 (m, 1H). 2.19-2.15 (m, 1H), 0.96 (d, J = 6.4 Hz, 6H).

EXAMPLE S79 Synthesis of (R) -3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (79)

Step 1 synthesis of (R) -3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S78 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS,21 x 250 mm,5 u column, flow rate of 1 mL/min, isocratic elution in 100% ACN (no additive) at 22 ℃. The title compound was isolated as the first eluting peak, rt=10.08 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) = 10.86 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.60 (d, J = 1.7 Hz, 1H), 7.57 - 7.50 (m, 2H), 7.00 (d, J = 1.2 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.86 (dd, J = 1.6, 8.9 Hz, 1H), 4.28 (dd, J = 5.1, 9.5 Hz, 1H), 3.91 (s, 5H), 3.82 - 3.73 (m, 2H), 3.67 (dd, J = 5.3, 12.8 Hz, 2H), 3.56 (s, 2H), 3.12 (s, 3H), 2.69 - 2.58 (m, 2H), 2.38 - 2.26 (m, 1H), 2.21 - 2.10 (m, 1H), 0.95 (d, J = 6.2 Hz, 6H).

EXAMPLE S80 Synthesis of (S) -3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (80)

Step 1 synthesis of (S) -3- (6- ((2R, 6R) -4- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2, 6-dimethylpiperazin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S78 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS,21 x 250 mm,5 u column, flow rate of 1 mL/min, isocratic elution in 100% ACN (no additive) at 22 ℃. The title compound was isolated as a second elution peak, rt=13.90 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) = 10.86 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.60 (d, J = 1.7 Hz, 1H), 7.57 - 7.50 (m, 2H), 7.00 (d, J = 1.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.86 (dd, J = 1.6, 8.9 Hz, 1H), 4.28 (dd, J = 5.2, 9.4 Hz, 1H), 3.91 (s, 5H), 3.82 - 3.74 (m, 2H), 3.67 (dd, J = 5.3, 12.7 Hz, 2H), 3.56 (s, 2H), 3.12 (s, 3H), 2.70 - 2.56 (m, 2H), 2.37 - 2.29 (m, 2H), 2.22 - 2.12 (m, 1H), 0.95 (d, J = 6.1 Hz, 6H).

EXAMPLE S81.3 Synthesis of 3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (81)

Step 1 (2S, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester synthesis. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (1.9G, 3.80 mmol), (2S, 4R) -4-amino-2-methyl-piperidine-1-carboxylic acid tert-butyl ester hydrochloride (1.05G, 4.18 mmol) and tBuXPhos-Pd-G3 (302 mg,0.380 mmol) and NaOtBu (912 mg,9.49 mmol) in MeTHF (38 mL) was evacuated and backfilled with nitrogen for 3 cycles. The mixture was heated to 85 ℃ for 1.5 h and cooled to room temperature. DCM (30 mL) was added and the mixture was filtered over a celite pad and rinsed with DCM (3×30 mL). The volatiles were concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-60% EtOAc in hexanes to give the title compound as a solid （2.01 g,84%）.MS (ESI) [M+H]⁺634.1.¹H NMR (400 MHz, DMSO-d₆) δ 7.87 (d, J = 8.1 Hz, 1H), 7.46 (d, J = 6.9 Hz, 2H), 7.42 – 7.21 (m, 9H), 6.55 (d, J = 8.1 Hz, 1H), 6.47 – 6.36 (m, 2H), 5.66 (d, J = 8.3 Hz, 1H), 5.44 (s, 2H), 5.41 (s, 2H), 4.45 – 4.33 (m, 1H), 3.89 (s, 3H), 3.88 – 3.86 (m, 1H), 3.76 – 3.62 (m, 1H), 3.01 (t, J = 13.9 Hz, 1H), 2.05 (d, J = 11.5 Hz, 1H), 1.87 – 1.83 (m, 1H), 1.48 – 1.43 (m, 1H), 1.41 (s, 9H), 1.23 (d, J= 7.0 Hz, 3H), 1.16 – 1.05 (m, 1H).

Step 2 Synthesis of 3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 4, 5 and 6 using (2 s,4 r) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.81 (s, 1H), 8.63 (s, 1H), 8.01 (s, 1H), 7.55 (s, 1H), 7.54 – 7.46 (m, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.93 (d, J = 8.4 Hz, 1H), 6.52 (dd, J = 8.8, 1.7 Hz, 1H), 6.47 (s, 1H), 5.67 (d, J = 8.3 Hz, 1H), 4.91 (br s, 1H), 4.48 (br s, 1H), 4.18 (dd, J = 8.8, 5.1 Hz, 1H), 3.83 (s, 3H), 3.81 – 3.74 (m, 1H), 3.54 (s, 2H), 3.11 (s, 3H), 3.06 (td, J = 13.1, 1.7 Hz, 1H), 2.66 – 2.56 (m, 2H), 2.31 – 2.22 (m, 1H), 2.17 – 2.08 (m, 2H), 1.92 (d, J = 11.3 Hz, 1H), 1.47 (td, J = 13.0, 5.7 Hz, 1H), 1.26 (d, J = 6.9 Hz, 3H), 1.15 (td, J = 13.9, 5.3 Hz, 1H).

EXAMPLE S82 Synthesis of (R) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (82)

Step 1 synthesis of (R) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S81 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% ACN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=11.30 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) 10.81 (s, 1H), 8.62 (s, 1H), 8.01 (s, 1H), 7.61 - 7.42 (m, 2H), 7.33 (d, J = 8.7 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.62 - 6.40 (m, 2H), 5.67 (br d, J = 8.5 Hz, 1H), 5.09 - 4.32 (m, 2H), 4.18 (dd, J = 5.1, 8.8 Hz, 1H), 3.91 - 3.69 (m, 4H), 3.54 (s, 2H), 3.11 (s, 4H), 2.31 - 2.06 (m, 3H), 1.91 (br d, J = 10.5 Hz, 1H), 1.46 (dt, J = 5.2, 11.9 Hz, 1H), 1.26 (br d, J = 6.7 Hz, 3H), 1.20 - 1.09 (m, 1H).

EXAMPLE S83 Synthesis of (S) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (83)

Step 2 synthesis of (S) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S81 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% ACN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt= 19.77 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) 10.81 (br s, 1H), 8.62 (br s, 1H), 8.01 (br s, 1H), 7.65 - 7.44 (m, 2H), 7.33 (br d, J = 8.8 Hz, 1H), 6.93 (br d, J = 8.1 Hz, 1H), 6.58 - 6.39 (m, 2H), 5.67 (br d, J = 8.5 Hz, 1H), 5.13 - 4.32 (m, 2H), 4.31 - 4.10 (m, 1H), 3.97 - 3.71 (m, 4H), 3.54 (br s, 2H), 3.11 (br s, 4H), 2.33 - 2.08 (m, 3H), 1.97 - 1.86 (m, 1H), 1.55 - 1.41 (m, 1H), 1.26 (br d, J = 6.6 Hz, 3H), 1.20 - 1.07 (m, 1H).

EXAMPLE S84.3 Synthesis of 3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (84)

Step 1 (2S, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester synthesis. A mixture of 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (1.9G, 3.80 mmol, intermediate provided by Celgene), (2S, 4R) -4-amino-2-methyl-piperidine-1-carboxylic acid tert-butyl ester hydrochloride (1.05G, 4.18 mmol) and tBuXPhos-Pd-G3 (302 mg,0.380 mmol) and NaOtBu (912 mg,9.49 mmol) in MeTHF (38 mL) was evacuated and backfilled with nitrogen for 3 cycles. The mixture was heated to 85 ℃ for 1.5 h and cooled to room temperature. DCM (30 mL) was added and the mixture was filtered over a celite pad and rinsed with DCM (3×30 mL). The volatiles were concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-60% EtOAc in hexanes to give the title compound as a solid （2.01 g,84%）.MS (ESI) [M+H]⁺ 634.1.¹H NMR (400 MHz, DMSO-d₆) δ 7.87 (d, J = 8.1 Hz, 1H), 7.46 (d, J = 6.9 Hz, 2H), 7.42 – 7.21 (m, 9H), 6.55 (d, J = 8.1 Hz, 1H), 6.47 – 6.36 (m, 2H), 5.66 (d, J = 8.3 Hz, 1H), 5.44 (s, 2H), 5.41 (s, 2H), 4.45 – 4.33 (m, 1H), 3.89 (s, 3H), 3.88 – 3.86 (m, 1H), 3.76 – 3.62 (m, 1H), 3.01 (t, J = 13.9 Hz, 1H), 2.05 (d, J = 11.5 Hz, 1H), 1.87 – 1.83 (m, 1H), 1.48 – 1.43 (m, 1H), 1.41 (s, 9H), 1.23 (d, J= 7.0 Hz, 3H), 1.16 – 1.05 (m, 1H).

Step 2 synthesis of (2S, 4R) -4- [ [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester. To a solution of (2 s,4 r) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester (525 mg,0.83 mmol) in DMSO (6.5 mL) was added sequentially formaldehyde solution (0.53 mL,7.09 mmol,37%) and decaborane (152 mg,1.24 mmol). The mixture was stirred at room temperature for 2 h. Water (25 mL) was added and the precipitated solid was collected by filtration. The residue was purified by column chromatography on silica gel using a gradient of 0-50% EtOAc in hexanes to give the title compound as a solid （449 mg,84%）.MS (ESI) [M+H]⁺ 648.7; ¹H NMR (500 MHz, DMSO-d₆) δ 7.89 (d, J = 8.1 Hz, 1H), 7.51 – 7.44 (m, 3H), 7.42 – 7.36 (m, 4H), 7.36 – 7.24 (m, 4H), 6.77 (dd, J = 9.3, 2.0 Hz, 1H), 6.63 (d, J = 1.8 Hz, 1H), 6.56 (d, J = 8.1 Hz, 1H), 5.48 – 5.41 (m, 2H), 5.41 (s, 2H), 4.51 – 4.36 (m, 1H), 4.10 (t, J = 11.9 Hz, 1H), 4.00 – 3.86 (m, 4H), 3.17 – 2.96 (m, 1H), 2.75 (s, 3H), 1.83 – 1.72 (m, 1H), 1.70 – 1.55 (m, 2H), 1.48 (d, J = 13.3 Hz, 1H), 1.41 (s, 9H), 1.25 – 1.19 (m, 3H).

Step 3 Synthesis of (2S, 4R) -4- [ [3- (2, 6-dioxo-3-piperidyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester. A mixture of (2 s,4 r) -4- [ [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester (446 mg,0.69 mmol) and peltier catalyst (96.7 mg,0.340 mmol) in THF (9 mL) and EtOH (9 mL) was stirred under a hydrogen atmosphere (1 atm) at 50 ℃ for 6 h and cooled to room temperature. The mixture was filtered over a pad of celite and rinsed with THF (3×30 mL). The volatiles were concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 10% -100% EtOAc in hexanes to give the title compound as a solid （303 mg,94%）.MS (ESI) [M+H]⁺470.3.¹H NMR (400 MHz, DMSO-d₆) δ 10.83 (s, 1H), 7.48 (d, J = 9.0 Hz, 1H), 6.88 (dd, J = 9.2, 1.9 Hz, 1H), 6.64 (d, J = 1.7 Hz, 1H), 4.49 – 4.36 (m, 1H), 4.24 (dd, J = 8.9, 5.1 Hz, 1H), 4.18 – 4.08 (m, 1H), 3.99 – 3.91 (m, 1H), 3.86 (s, 3H), 3.15 – 2.97 (m, 1H), 2.75 (s, 3H), 2.68 – 2.55 (m, 2H), 2.33 – 2.23 (m, 1H), 2.22 – 2.12 (m, 1H), 1.84 – 1.71 (m, 1H), 1.71 – 1.55 (m, 2H), 1.49 (d, J = 12.1 Hz, 1H), 1.41 (s, 9H), 1.25 – 1.17 (m, 3H).

Step 4 Synthesis of 3- [ 1-methyl-6- [ methyl- [ (2S, 4R) -2-methyl-4-piperidinyl ] amino ] indazol-3-yl ] piperidine-2, 6-dione hydrochloride. To a solution of (2 s,4 r) -4- [ [3- (2, 6-dioxo-3-piperidinyl) -1-methyl-indazol-6-yl ] -methyl-amino ] -2-methyl-piperidine-1-carboxylic acid tert-butyl ester (400 mg,0.85 mmol) in 1, 4-dioxane (5 mL) was added 4N HCl (2.13 mL,8.52 mmol) in 1, 4-dioxane, and the mixture was stirred at room temperature for 15 h. The volatiles were evaporated under reduced pressure and the residue was triturated in Et ₂ O (10 mL) to give the title compound as a solid (295 mg, 85%) which was used in the next step without further purification .MS (ESI) [M+H]⁺370.3.¹H NMR (500 MHz, DMSO-d₆) δ 10.88 (s, 1H), 9.12 (br s, 1H), 9.05 – 8.83 (m, 1H), 7.71 (br s, 1H), 7.21 (br s, 1H), 4.40 – 4.30 (m, 1H), 4.27 – 3.98 (m, 6H), 3.95 (s, 3H), 3.89 – 3.73 (m, 1H), 3.34 – 3.24 (m, 1H), 3.22 – 3.11 (m, 1H), 3.07 – 2.85 (m, 2H), 2.75 – 2.55 (m, 2H), 2.42 – 2.29 (m, 1H), 2.19 – 2.11 (m, 1H), 1.35 (d, J = 5.4 Hz, 3H).

Step 5 Synthesis of 3- [6- [ [ (2S, 4R) -1- [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -2-methyl-4-piperidinyl ] -methyl-amino ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione. To a solution of 3- [ 1-methyl-6- [ methyl- [ (2 s,4 r) -2-methyl-4-piperidinyl ] amino ] indazol-3-yl ] piperidine-2, 6-dione hydrochloride (64.1 mg,0.16 mmol) in DMF (2 mL) and DMSO (0.5 mL) was added sequentially 5- [ (5-chloro-2-fluoro-pyrimidin-4-yl) amino ] -1-methyl-indolin-2-one (42 mg,0.14 mmol) and DIPEA (0.2 ml,1.15 mmol). The mixture was heated to 100 ℃ for 18 h and then cooled to room temperature. Volatiles were removed under reduced pressure and the residue was purified by using a gradient of 20% -75% MeCN and 10mM ammonium formate in water to give the title compound as a solid (47 mg, 48%). LCMS C ₃₃H₃₆ClN₉O₃ theory 641.3, found 642.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.83 (s, 1H), 8.63 (s, 1H), 8.01 (s, 1H), 7.56 (br s, 1H), 7.52 – 7.43 (m, 2H), 6.96 – 6.85 (m, 2H), 6.65 (d, J = 1.6 Hz, 1H), 5.17 – 4.78 (m, 1H), 4.72 – 4.36 (m, 1H), 4.29 – 4.16 (m, 2H), 3.87 (s, 3H), 3.54 (s, 2H), 3.15 – 3.03 (m, 4H), 2.75 (s, 3H), 2.68 – 2.54 (m, 2H), 2.34 – 2.26 (m, 1H), 2.17 (td, J = 11.9, 6.0 Hz, 1H), 1.86 – 1.71 (m, 2H), 1.71 – 1.52 (m, 2H), 1.25 (d, J = 6.8 Hz, 3H).

EXAMPLE S85.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (85)

Step 1 synthesis of 6-bromo-1-methyl-indazole-3-carbaldehyde cesium carbonate (14.480 g,44.44 mmol,2.00 eq) and methyl iodide (3.780 g,26.66 mmol,1.7 mL,1.20 eq) were added to a solution of 6-bromo-1H-indazole-3-carbaldehyde (5.000 g,22.22 mmol,1.00 eq) in dimethylformamide (50 mL) and the mixture was stirred at 50 ℃ for 12H. The reaction mixture was cooled to room temperature. The mixture was filtered and the filtrate was added to ethyl acetate (100 mL) and water (50 mL) and the organic layer was separated. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over magnesium sulfate, filtered and the filtrate was concentrated to give the product. The residue was purified by column on silica gel (5% -10% ethyl acetate in petroleum ether). The title compound was obtained as a white solid (3.500 g,14.64 mmol,65.9% yield). LCMS 240.9 [ M+H ] ⁺.

Step 2 (synthesis of 6-bromo-1-methyl-indazol-3-yl) methanol sodium borohydride (0.886 g,23.42 mmol,2.00 eq) was added to a solution of 6-bromo-1-methyl-indazol-3-carbaldehyde (2.800 g,11.71 mmol,1.00 eq) in ethanol (40 mL) at 0 ℃. The mixture was stirred at 20 ℃ for 1 h. The 6-bromo-1-methyl-indazole-3-carbaldehyde was consumed and the desired mass was detected by LCMS. The mixture was filtered and the filtrate was added to ethyl acetate (50 mL) and water (50 mL) under pressure and the organic layer was separated. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over magnesium sulfate, filtered and the filtrate concentrated to give the title compound (3.000 g, crude) as a white solid. LCMS 242.0 [ M+H ] ⁺.

Step 3 Synthesis of 6-bromo-3- (chloromethyl) -1-methyl-1H-indazole to a solution of (6-bromo-1-methyl-indazol-3-yl) methanol (3.000 g,12.44 mmol,1.00 eq) in dichloromethane (50 mL) was added thionyl chloride (2.960 g,24.89 mmol,1.8 mL,2.00 eq) at 0 ℃. The mixture was stirred at 20 ℃ for 2 h. (6-bromo-1-methyl-indazol-3-yl) methanol was consumed and the desired mass was detected by LCMS. The mixture was concentrated under pressure. The mixture was filtered, and the filtrate was concentrated. The residue was purified by column on silica gel (5% -20% ethyl acetate in petroleum ether). The title compound was obtained as a yellow solid (1.600 g,6.16 mmol,49.5% yield). LCMS 260.9[ m+1] +. ¹ H NMR (400 MHz, chloroform -d) δ, 7.69 (d, J =8.8 Hz, 1H), 7.59 (s, 1H), 7.31 (m, J =1.2, 8.4 Hz, 1H), 4.93 (s, 2H), 4.02 (s, 3H))

Step 4 Synthesis of 2- (6-bromo-1-methyl-1H-indazol-3-yl) acetonitrile to a solution of 6-bromo-3- (chloromethyl) -1-methyl-1H-indazol (1.95 g,7.51 mmol) in DMF (25.05 ml) was added sodium cyanide (0.479 g,9.77 mmol) and the mixture stirred at 60℃for 3H. The reaction mixture was cooled to room temperature, and ethyl acetate (100 mL) and water (50 mL) were added. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined extracts were washed with saturated brine (50 mL), dried over magnesium sulfate, filtered and concentrated to give a yellow solid (1.867 g,7.47 mmol,99% yield). As such, was used in the following steps. LCMS 249.8 [ M+H ] ⁺.

Step 5 Synthesis of 2- (6-bromo-1-methyl-1H-indazol-3-yl) propionitrile to a solution of 2- (6-bromo-1-methyl-1H-indazol-3-yl) acetonitrile (1.867 g,7.47 mmol) in tetrahydrofuran (46.7 ml) was added potassium tert-butoxide (1.005 g,8.96 mmol) at-20℃and after stirring the mixture at-20℃for 30min, a solution of methyl iodide (0.467 ml,7.47 mmol) was added dropwise. The mixture was stirred overnight while warming to room temperature. The mixture was filtered, and the filtrate was concentrated. The residue was purified with 20% EtOAc in hexanes and the title compound was obtained as a yellow oil (0.5885 g,2.228 mmol,29.8% yield). LCMS 265.1 [ M+H ] ⁺.¹ H NMR (chloroform -d, 400 MHz) δ 7.6-7.8 (m, 1H), 7.5-7.6 (m, 1H), 7.3-7.5 (m, 1H), 4.3-4.4 (m, 1H), 4.39 (s, 1H), 3.9-4.1 (m, 5H), 1.6-1.9 (m, 3H)

Step 6 Synthesis of tert-butyl 4- (6-bromo-1-methyl-1H-indazol-3-yl) -4-cyanovalerate to a solution of 2- (6-bromo-1-methyl-1H-indazol-3-yl) propionitrile (0.300 g,1.136 mmol) and tert-butyl acrylate (0.333 ml,2.272 mmol) in toluene (4.54 ml) was added potassium carbonate (0.314 g,2.272 mmol) and N-benzyl-N, N-diethylethylammonium chloride (0.026 g,0.114 mmol). The mixture was stirred at 65 ℃ for 12 h. After cooling, the mixture was filtered and the filtrate was concentrated. The residue was further purified by preparative TLC (20% ethyl acetate in hexane). Obtained as a yellow oil (0.212 g,0.540 mmol,47.6% yield). LCMS 336.1 (M- ^tBu+H)⁺.¹ H NMR (chloroform -d, 400 MHz) δ 7.84 (s, 1H), 7.8-7.9 (m, 1H), 7.57 (s, 1H), 7.5-7.7 (m, 1H), 7.30 (s, 1H), 7.3-7.3 (m, 1H), 4.00 (s, 3H), 2.3-2.6 (m, 4H), 1.87 (s, 3H), 1.41 (s, 9H)

Step 7 Synthesis of (3R, 4R) -4- ((3- (5- (tert-butoxy) -2-cyano-5-oxopent-2-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl 4- (6-bromo-1-methyl-1H-indazol-3-yl) -4-cyanovalerate (25 mg,0.064 mmol), bis (dibenzylideneacetone) palladium (0) (0.366 mg, 0.637. Mu. Mol), sodium tert-butoxide (12.25 mg,0.127 mmol), 2 '-bis (diphenylphosphino) -1,1' -binaphthyl (1.190 mg, 1.912. Mu. Mol), (3R, 4R) -4-amino-3-methylpiperidine-1-carboxylic acid tert-butyl ester (20.49 mg,0.096 mmol) and PhMe (319. Mu.L) were charged into a microwave vial. It was purged with N ₂ for five minutes and then heated at 80 ℃ overnight. It was filtered through a celite plug, rinsing with EtOAc and DCM to give the crude product. It was purified via normal phase chromatography with MeOH in DCM. The title compound was obtained as a yellow solid. LCMS 470.0 [ M+H ] ⁺.¹ H NMR (chloroform -d, 400 MHz) δ 7.5-7.8 (m, 1H), 6.1-6.6 (m, 2H), 4.0-4.2 (m, 3H), 3.7-3.9 (m, 3H), 3.0-3.2 (m, 2H), 2.8-3.0 (m, 3H), 2.3-2.7 (br m, 2H), 2.0-2.2 (m, 3H), 1.6-1.8 (m, 3H), 1.47 (s, 11H), 1.2-1.3 (m, 8H), 0.9-1.1 (m, 3H), 0.8-0.9 (m, 2H), 0.8-0.9 (m, 2H)

Step 8 Synthesis of 5-amino-4- (6- (((3R, 4R) -1- (tert-butoxycarbonyl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -4-methyl-5-oxopentanoic acid to a mixture of tert-butyl (3R, 4R) -4- ((3- (5- (tert-butoxy) -2-cyano-5-oxopent-2-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidine-1-carboxylate (32.3 mg,0.061 mmol) in DMSO (307. Mu.L) and ethanol (307. Mu.L) was added sodium hydroxide (123. Mu.L, 0.123 mmol) and hydrogen peroxide (126. Mu.L, 1.229 mmol) at 0 ℃. The reaction mixture was stirred at 20 ℃ for 12 h.4- (6-bromo-1-methyl-1H-indazol-3-yl) -4-cyanopentanoic acid tert-butyl ester was consumed and the desired mass was detected by LCMS. The mixture was quenched with saturated aqueous ammonium chloride. The pH was adjusted to 5 with 1M hydrochloric acid and the mixture was extracted with ethyl acetate (30 mL x 2), then the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the title compound (30 mg,0.062 mmol,100% yield) as a white solid. LCMS 488.20 [ M+H ] ⁺.¹ H NMR (chloroform -d, 400 MHz) δ 7.4-7.5 (m, 1H), 6.4-6.5 (m, 1H), 6.2-6.3 (m, 1H), 5.9-6.0 (m, 1H), 4.6-4.9 (m, 2H), 3.9-4.0 (m, 3H), 3.1-3.2 (m, 2H), 2.7-2.9 (m, 3H), 2.2-2.5 (m, 2H), 2.1-2.2 (m, 1H), 2.0-2.1 (m, 1H), 1.5-1.6 (m, 2H), 1.47 (s, 9H), 1.2-1.4 (m, 7H), 1.0-1.1 (m, 3H)

Step 9 Synthesis of tert-butyl (3R, 4R) -3-methyl-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidine-1-carboxylate CDI (29.9 mg,0.185 mmol) and DMAP (0.752 mg, 6.15. Mu. Mol) were added to a solution of 5-amino-4- (6- (((3R, 4R) -1- (tert-butoxycarbonyl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -4-methyl-5-oxopentanoic acid (30 mg,0.062 mmol) in THF (615. Mu.L) and the mixture was stirred at 70℃for 12H. The mixture was filtered and the filtrate was concentrated. It was purified via reverse phase (accq prep) with a gradient of 0-100% acetonitrile in water. The fractions were concentrated to give the title compound as a white solid (21.89 mg,0.047 mmol,76% yield). LCMS 470.0 [ M+H ] ⁺.¹ H NMR (chloroform -d, 400 MHz) δ 7.86 (s, 1H), 7.8-8.1 (m, 1H), 7.5-7.7 (m, 1H), 6.4-6.6 (m, 1H), 6.2-6.3 (m, 1H), 4.0-4.2 (m, 2H), 3.87 (s, 3H), 3.0-3.2 (m, 1H), 2.5-3.0 (m, 5H), 2.0-2.3 (m, 2H), 2.01 (s, 3H), 1.6-1.7 (m, 2H), 1.47 (s, 9H), 1.2-1.3 (m, 2H), 1.0-1.1 (m, 3H)

Step 10 Synthesis of 3-methyl-3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione A2-part vial was charged with (3R, 4R) -3-methyl-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidine-1-carboxylic acid tert-butyl ester (21.89 mg,0.047 mmol) in DCM (466. Mu.L) and hydrochloric acid (4M in dioxane) (58.3. Mu.L, 0.233 mmol) was added dropwise. It was stirred at room temperature for 1 hour. Thereafter, the solution was concentrated to give the title compound (17.22 mg,0.047 mmol,100% yield) as a white solid. As such for the next step. LCMS 370.5 [ M+H ] ⁺.

Step 11 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidin-2, 6-dione A solution of 3-methyl-3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione (17 mg,0.046 mmol), 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (13.47 mg,0.046 mmol), DIPEA (0.020 mL,0.115 mmol) in DMSO (0.5 mL) was stirred at 80℃for 2 hours. After that, the reaction was quenched with 10% formic acid in DMSO and filtered through a syringe filter. The crude material was purified by preparative HPLC to give the title compound as an off-white solid (30 mg,0.045 mmol,23% yield). LCMS C ₃₁H₃₀ClFN₁₀O₃ theory 642.2, found 642.8 [ m+h ] ⁺;¹ H NMR (chloroform -d, 400 MHz) δ 7.98 (s, 1H), 7.76 (s, 1H), 7.7-7.8 (m, 1H), 7.5-7.6 (m, 2H), 7.4-7.5 (m, 1H), 6.9-7.1 (m, 1H), 6.7-6.8 (m, 1H), 6.4-6.6 (m, 1H), 6.2-6.3 (m, 1H), 4.5-4.7 (m, 2H), 3.88 (s, 3H), 3.6-3.8 (m, 1H), 3.4-3.6 (m, 2H), 3.21 (s, 3H), 3.0-3.1 (m, 1H), 2.8-2.9 (m, 1H), 2.7-2.8 (m, 2H), 2.5-2.7 (m, 1H), 2.2-2.3 (m, 1H), 2.0-2.1 (m, 1H), 1.75 (s, 3H), 1.6-1.7 (m, 1H), 1.2-1.4 (m, 1H), 1.0-1.1 (m, 3H)

EXAMPLE S86.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [4,3-c ] pyridin-3-yl) piperidine-2, 6-dione (86)

Step 1.6-chloro-3-iodo-1-methyl-1H-pyrazolo [4,3-c ] pyridine sodium hydride (0.150 g,3.76 mmol) was added to a 0 ℃ solution of 6-chloro-3-iodo-1H-pyrazolo [4,3-c ] pyridine (1 g,3.58 mmol) in DMF (20 mL), and the reaction mixture was stirred for 30 min. Sodium hydride (0.150 g,3.76 mmol) was then added dropwise. The reaction mixture was allowed to warm to room temperature and stirred again 1 h. The reaction mixture was quenched with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound as a white solid (0.75 g,2.56 mmol,71.4% yield ）.MS (ESI) m/z 293.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.59 (s, 1 H), 7.92 (s, 1 H), 4.06 (d, J=0.98 Hz, 3 H).

Step 2.3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-chloro-1-methyl-1H-pyrazolo [4,3-c ] pyridine A solution of 6-chloro-3-iodo-1-methyl-1H-pyrazolo [4,3-c ] pyridine (415 mg,1.414 mmol), (2, 6-bis (benzyloxy) pyridin-3-yl) boronic acid (521 mg,1.555 mmol) and sodium bicarbonate (261 mg,3.11 mmol) in a mixed solvent of DME (10 mL) and water (2 mL) was stirred with tetrakis (triphenylphosphine) palladium (0) (163 mg,0.141 mmol) at 100℃for 15H. The reaction mixture was treated with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give the title compound as a white solid (300 mg,0.657 mmol,46.4% yield ）.MS (ESI) m/z 457.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.89 (d, J=0.98 Hz, 1 H), 8.02 (d, J=8.19 Hz, 1 H), 7.86 (d, J=0.98 Hz, 1 H), 7.26 - 7.49 (m, 10 H), 6.63 (d, J=8.19 Hz, 1 H), 5.52 (s, 2 H), 5.44 (s, 2 H), 4.02 - 4.08 (m, 3 H).

3. Tert-butyl (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [4,3-c ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylate to a solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-chloro-1-methyl-1H-pyrazolo [4,3-c ] pyridine (400 mg,0.875 mmol), ruPhos Pd G (73.2 mg,0.088 mmol) and tert-butyl (3R, 4R) -4-amino-3-methylpiperidine-1-carboxylate (188 mg,0.875 mmol) in 1, 4-dioxane (5 mL) was added cesium carbonate (570 mg,1.751 mmol). The reaction mixture was purged with argon 1 min and they were stirred at 100 ℃ 15: 15 h. LCMS indicated completion of the reaction. The reaction mixture was treated with saturated aqueous sodium chloride (25 mL) and then extracted with ethyl acetate (3X 25 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (1X 25 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered through celite and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (0-100% ethyl acetate in hexane). The desired fractions were concentrated under reduced pressure to give a solid which was further purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% TFA in water (0.1% TFA), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (95 mg,0.150 mmol,17.1% yield ）.MS (ESI) m/z 635.0 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.60 (br s, 1 H), 7.96 (d, J=8.07 Hz, 1 H), 7.23 - 7.51 (m, 10 H), 6.59 (d, J=8.07 Hz, 1 H), 6.34 (br s, 1 H), 5.49 (s, 2 H), 5.41 (s, 2 H), 3.93 (br s, 2 H), 3.87 (s, 3 H), 3.44 - 3.56 (m, 1 H), 2.88 (br s, 1 H), 2.51 - 2.62 (m, 1 H), 1.92 (br d, J=9.66 Hz, 1 H), 1.52 - 1.63 (m, 1 H), 1.41 (s, 9 H), 1.16 - 1.29 (m, 1 H), 0.89 (d, J=6.48 Hz, 3 H).

Step 4.3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-pyrazolo [4,3-c ] pyridin-3-yl) piperidine-2, 6-dione A solution of (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-pyrazolo [4,3-c ] pyridin-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester (0.095 g,0.150 mmol) in ethanol (10 mL) was stirred under 80℃with 10% w/w (0.016 g,0.015 mmol) palladium on carbon 15H. The reaction mixture was filtered through celite, the eluate was concentrated and then the residue was stirred with 1 mL TFA in DCM (1 mL) at room temperature for 30 min. TFA and solvent were removed under reduced pressure to give the desired product as TFA salt, which was used without further purification (53 mg,0.149 mmol,99% yield).

Step 5.3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-pyrazolo [4,3-c ] pyridin-3-yl) piperidine-2, 6-dione the title compound was synthesized using general procedure 6 using 3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-pyrazolo [4,3-c ] pyridin-3-yl) piperidine-2, 6-dione (53 mg,0.149 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (43.5 mg,0.149 mmol). The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (7.7 mg,0.012 mmol,8.23% yield ）.MS (ESI) m/z 628.8 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.89 (s, 1 H), 8.65 (s, 1 H), 8.49 (d, J=0.86 Hz, 1 H), 8.01 (s, 1 H), 7.57 (s, 1 H), 7.52 (dd, J=8.50, 1.90 Hz, 1 H), 6.94 (d, J=8.44 Hz, 1 H), 6.23 - 6.31 (m, 2 H), 4.42 - 4.59 (m, 2 H), 4.28 (dd, J=9.84, 5.07 Hz, 1 H), 3.78 (s, 3 H), 3.55 - 3.66 (m, 1 H), 3.53 (s, 2 H), 3.10 (s, 3 H), 2.89 - 3.04 (m, 1 H), 2.54 - 2.65 (m, 2 H), 2.35 - 2.41 (m, 1 H), 2.09 - 2.23 (m, 1 H), 1.98 (br d, J=10.39 Hz, 1 H), 1.54 - 1.72 (m, 1 H), 1.16 - 1.37 (m, 1 H), 0.93 (d, J=6.36 Hz, 3 H).

EXAMPLE S87 Synthesis of (R) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (87)

Step 1 synthesis of (R) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S84 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% ACN (no additives)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=13.41 min. LCMS C ₃₃H₃₆ClN₉O₃ theory 641.3, found 642.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ (ppm) 10.84 (s, 1H), 8.64 (s, 1H), 8.01 (s, 1H), 7.56 (br s, 1H), 7.52 - 7.45 (m, 2H), 6.96 - 6.88 (m, 2H), 6.65 (d, J = 1.6 Hz, 1H), 4.24 (dd, J = 8.9, 5.1 Hz, 2H), 3.87 (s, 3H), 3.54 (s, 2H), 3.15 - 3.04 (m, 4H), 2.75 (s, 3H), 2.69 - 2.58 (m, 2H), 2.37 - 2.06 (m, 2H), 1.86 - 1.52 (m, 4H), 1.25 (br d, J = 6.8 Hz, 3H).

EXAMPLE S88 Synthesis of (S) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (88)

Step 1 synthesis of (S) -3- (6- (((2S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -2-methylpiperidin-4-yl) (methyl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S84 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1mL/min flow rate, isocratic elution in 100% MECN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt=23.26 min. LCMS C ₃₃H₃₆ClN₉O₃ theory 641.3, found 642.3 [M+H]⁺;¹H NMR (400 MHz, CD₃SOCD₃, 298 K) δ (ppm) = 10.84 (s, 1H), 8.64 (s, 1H), 8.01 (s, 1H), 7.56 (br s, 1H), 7.53 - 7.44 (m, 2H), 6.96 - 6.87 (m, 2H), 6.65 (d, J = 1.6 Hz, 1H), 4.24 (dd, J = 9.1, 5.1 Hz, 2H), 3.87 (s, 3H), 3.54 (s, 2H),3.15 - 3.04 (m, 4H), 2.75 (s, 3H), 2.68 - 2.58 (m, 2H), 2.36 - 2.09 (m, 2H), 1.88 - 1.51 (m, 4H), 1.24 (br d, J = 7.0 Hz, 3H).

EXAMPLE S89.3 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (89)

Step 1 Synthesis of 3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1-ethylindol-2-one and 3- (1-methyl-6- (piperidin-4-ylamino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 8.64 (s, 1H), 8.44 (br, 1H), 8.01 (s, 1H), 7.54 (s, 1H), 7.54 – 7.49 (m, 1H), 7.32 (d, J= 8.7 Hz, 1H), 6.98 (d, J= 8.4 Hz, 1H), 6.52 (dd, J= 8.8, 1.7 Hz, 1H), 6.46 (s, 1H), 5.82 – 5.73 (m, 1H), 4.42 – 4.35 (m, 2H), 4.18 (dd, J= 8.8, 5.1 Hz, 1H), 3.82 (s, 3H), 3.67 (q, J= 7.2 Hz, 2H), 3.53 (s, 2H), 3.11 (t, J= 11.3 Hz, 2H), 2.64 – 2.56 (m, 2H), 2.30 – 2.21 (m, 1H), 2.18 – 2.10 (m, 1H), 2.04 – 1.94 (m, 2H), 1.36 – 1.26 (m, 2H), 1.13 (t, J= 7.1 Hz, 3H).

EXAMPLE S90 Synthesis of (R) -3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (90)

Step 1 (R) -Synthesis of 3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S89 was then isolated using preparative HPLC (having REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MECN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=11.16 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 10.81 (s, 1H), 8.63 (s, 1H), 8.01 (s, 1H), 7.60 - 7.47 (m, 2H), 7.32 (d, J = 8.8 Hz, 1H), 6.98 (d, J = 8.5 Hz, 1H), 6.52 (dd, J = 1.5, 8.8 Hz, 1H), 6.46 (s, 1H), 5.76 (d, J = 8.2 Hz, 1H), 4.38 (br d, J = 13.4 Hz, 2H), 4.18 (dd, J = 5.1, 8.8 Hz, 1H), 3.82 (s, 3H), 3.71 - 3.59 (m, 3H), 3.53 (s, 2H), 3.11 (br t, J = 11.0 Hz, 2H), 2.62 - 2.58 (m, 2H), 2.30 - 2.21 (m, 1H), 2.19 - 2.09 (m, 1H), 2.00 (br d, J = 9.4 Hz, 2H), 1.38 - 1.26 (m, 2H), 1.13 (t, J = 7.2 Hz, 3H)

EXAMPLE S91 Synthesis of (S) -3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (91)

Step 1 (S) -Synthesis of 3- (6- ((1- (5-chloro-4- ((1-ethyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S89 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt=18.20 min. LCMS C ₃₂H₃₄ClN₉O₃ theory 627.3, found 628.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 10.81 (s, 1H), 8.63 (s, 1H), 8.02 (s, 1H), 7.58 - 7.50 (m, 2H), 7.32 (d, J = 8.8 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 6.55 - 6.49 (m, 1H), 6.46 (s, 1H), 5.76 (d, J = 8.3 Hz, 1H), 4.43 - 4.35 (m, 2H), 4.18 (dd, J = 5.2, 8.9 Hz, 1H), 3.82 (s, 3H), 3.71 - 3.59 (m, 3H), 3.53 (s, 2H), 3.11 (br t, J = 11.2 Hz, 2H), 2.62 - 2.58 (m, 2H), 2.32 - 2.20 (m, 1H), 2.19 - 2.11 (m, 1H), 2.00 (br d, J = 8.6 Hz, 2H), 1.37 - 1.27 (m, 2H), 1.13 (t, J = 7.2 Hz, 3H).

EXAMPLE S92.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (92)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-6-fluoro-1-methylindolin-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm 10.81 (s, 1H), 8.58 (s, 1H), 8.00 (s, 1H), 7.27 - 7.33 (m, 2H), 7.02 (d, J = 10.64 Hz, 1H), 6.49 (dd, J = 8.80, 1.59 Hz, 1H), 6.40 (s, 1H), 5.67 (d, J = 9.05 Hz, 1H), 4.33 (br s, 2H), 4.16 (dd, J = 8.74, 5.32 Hz, 1H), 3.80 (s, 3H), 3.53 (s, 2H), 3.23 (br d, J = 5.38 Hz, H), 3.10 (s, 3 H), 2.91 (br t, J = 11.74 Hz, 1H), 2.54 - 2.64 (m, 3H), 2.19 - 2.30 (m, 1H), 2.09 - 2.19 (m, 1H), 1.99 (br d, J = 10.15 Hz, 1H), 1.45 - 1.56 (m, 1H), 1.10 (br d, J = 13.33 Hz, 1H), 0.90 (br d, J = 6.48 Hz, 3H).

EXAMPLE S93 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (93)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S92 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=8.86 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm 10.80 (s, 1H), 8.57 (s, 1H), 8.00 (s, 1H), 7.36 - 7.27 (m, 2H), 7.02 (d, J = 10.6 Hz, 1H), 6.49 (dd, J = 1.5, 8.8 Hz, 1H), 6.40 (s, 1H), 5.66 (d, J = 9.1 Hz, 1H), 4.42 - 4.25 (m, 2H), 4.16 (dd, J = 5.1, 8.8 Hz, 1H), 3.80 (s, 3H), 3.53 (s, 2H), 3.23 (dq, J = 3.9, 9.7 Hz, 1H), 3.10 (s, 3H), 2.96 - 2.87 (m, 1H), 2.64 - 2.57 (m, 3H), 2.31 - 2.19 (m, 1H), 2.18 - 2.09 (m, 1H), 1.99 (br d, J = 10.0 Hz, 1H), 1.58 - 1.43 (m, 1H), 1.17 - 1.04 (m, 1H), 0.90 (br d, J = 6.3 Hz, 3H).

EXAMPLE S94 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (94)

Step 1 synthesis of(s) -3- (6- (((3 r,4 r) -1- (5-chloro-4- ((6-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S92 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt=12.57 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm10.80 (s, 1H), 8.57 (s, 1H), 8.00 (s, 1H), 7.35 - 7.27 (m, 2H), 7.02 (d, J = 10.6 Hz, 1H), 6.49 (dd, J = 1.7, 8.8 Hz, 1H), 6.40 (s, 1H), 5.66 (d, J = 9.1 Hz, 1H), 4.45 - 4.23 (m, 2H), 4.16 (dd, J = 5.2, 8.8 Hz, 1H), 3.80 (s, 3H), 3.53 (s, 2H), 3.27 - 3.17 (m, 1H), 3.10 (s, 3H), 2.97 - 2.86 (m, 1H), 2.61 - 2.57 (m, 3H), 2.29 - 2.19 (m, 1H), 2.18 - 2.09 (m, 1H), 1.99 (br d, J = 9.9 Hz, 1H), 1.55 - 1.47 (m, 1H), 1.17 - 1.04 (m, 1H), 0.90 (br d, J = 6.3 Hz, 3H).

EXAMPLE S95.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (95)

Step 1 synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the title compound was synthesized according to general procedures 1 and 6 using 5-amino-7-fluoro-1-methylindolin-2-one and 3- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm 10.81 (s, 1H), 8.79 (s, 1H), 8.06 (s, 1H), 7.61 (br d, J = 13.57 Hz, 1H), 7.43 (s, 1 H), 7.31 (d, J = 8.80 Hz, 1H), 6.41 - 6.61 (m, 2H), 5.70 (d, J = 8.93 Hz, 1H), 4.40 - 4.59 (m, 2H), 4.17 (dd, J = 8.68, 5.26 Hz, 1H), 3.81 (s, 3H), 3.61 (s, 2H), 3.26 (d, J = 2.45 Hz, H), 3.01 - 3.11 (m, 1H), 2.69 - 2.78 (m, 1H), 2.56 - 2.64 (m, 2H), 2.19 - 2.30 (m, 1H), 2.04 - 2.18 (m, 2H), 1.53 - 1.67 (m, 1H), 1.11 - 1.26 (m, 1H), 0.99 (d, J = 6.48 Hz, 3H).

EXAMPLE S96 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (96)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S95 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=10.48 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm 10.81 (s, 1H), 8.79 (s, 1H), 8.06 (s, 1H), 7.62 (br d, J = 13.1 Hz, 1H), 7.44 (br s, 1H), 7.31 (br d, J = 8.6 Hz, 1H), 6.51 (br d, J = 8.8 Hz, 1H), 6.45 (s, 1H), 5.70 (br d, J = 8.0 Hz, 1H), 4.58 - 4.41 (m, 2H), 4.25 - 4.12 (m, 1H), 3.82 (br s, 3H), 3.62 (br s, 2H), 3.27 (br s, 3H), 3.14 - 3.02 (m, 1H), 2.83 - 2.69 (m, 1H), 2.64 - 2.59 (m, 1H), 2.33 - 2.04 (m, 3H), 1.69 - 1.55 (m, 1H), 1.31 - 1.13 (m, 1H), 0.99 (br d, J = 5.6 Hz, 3H)

EXAMPLE S97 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (97)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((7-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the product of example S95 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, flow rate of 1mL/min, isocratic elution in 100% MeCN (no additive) at 22 ℃. The title compound was isolated as a second elution peak, rt=15.07 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ ppm10.81 (s, 1H), 8.79 (s, 1H), 8.07 (s, 1H), 7.62 (br d, J = 14.2 Hz, 1H), 7.44 (br s, 1H), 7.32 (br d, J = 8.7 Hz, 1H), 6.52 (br d, J = 8.9 Hz, 1H), 6.45 (s, 1H), 5.70 (br d, J = 8.7 Hz, 1H), 4.49 (br t, J = 13.4 Hz, 2H), 4.18 (dd, J = 5.3, 8.4 Hz, 1H), 3.82 (s, 3H), 3.62 (s, 2H), 3.27 (br s, 3H), 3.07 (t, J = 12.2 Hz, 1H), 2.82 - 2.69 (m, 1H), 2.66 - 2.57 (m, 2H), 2.31 - 2.08 (m, 3H), 1.66 - 1.57 (m, 1H), 1.29 - 1.13 (m, 1H), 1.00 (d, J = 6.3 Hz, 3H).

EXAMPLE S98.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (98)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-4-fluoro-1-methylindolin-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ ppm 10.81 (s, 1 H), 8.64 (s, 1 H), 8.01 (s, 1 H), 7.35 (t, J=7.95 Hz, 1 H), 7.29 (d, J=8.80 Hz, 1 H), 6.84 (d, J=8.31 Hz, 1 H), 6.49 (dd, J=8.86, 1.53 Hz, 1 H), 6.40 (s, 1 H), 5.67 (d, J=8.93 Hz, 1 H), 4.21 - 4.48 (m, 2 H), 4.12 - 4.21 (m, 1 H), 3.79 (s, 3 H), 3.53 - 3.70 (m, 2 H), 3.23 (qd, J=9.78, 3.79 Hz, 1 H), 3.13 (s, 3 H), 2.86 - 2.98 (m, 1 H), 2.52 - 2.63 (m, 3 H), 2.19 - 2.31 (m, 1 H), 2.08 - 2.19 (m, 1 H), 1.93 - 2.05 (m, 1 H), 1.43 - 1.60 (m, 1 H), 0.99 - 1.17 (m, 1 H), 0.79 - 0.96 (m, 3 H).

EXAMPLE S99 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (99)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S98 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=8.56 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ ppm 10.80 (br s, 1H), 9.99 - 9.96 (m, 1H), 8.63 (br s, 1H), 8.00 (s, 1H), 7.39 - 7.26 (m, 2H), 6.84 (br d, J = 8.1 Hz, 1H), 6.49 (br d, J = 8.5 Hz, 1H), 6.40 (br s, 1H), 5.66 (br d, J = 8.8 Hz, 1H), 4.45 - 4.22 (m, 2H), 4.16 (br dd, J = 4.9, 8.5 Hz, 1H), 3.79 (s, 3H), 3.70 - 3.51 (m, 2H), 3.26 - 3.18 (m, 1H), 3.12 (s, 3H), 2.96 - 2.87 (m, 1H), 2.67 - 2.55 (m, 3H), 2.29 - 2.19 (m, 1H), 2.18 - 2.10 (m, 1H), 1.99 (br d, J = 10.1 Hz, 1H), 1.54 - 1.43 (m, 1H), 1.15 - 1.04 (m, 1H), 0.89 (br s, 3H).

EXAMPLE S100 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (100)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((4-fluoro-1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S98 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt=12.25 min. LCMS C ₃₂H₃₃ClFN₉O₃ theory 645.2, found 646.4 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ ppm 10.80 (s, 1H), 8.63 (br s, 1H), 8.00 (s, 1H), 7.46 - 7.20 (m, 2H), 6.84 (br d, J = 8.2 Hz, 1H), 6.49 (br d, J = 8.8 Hz, 1H), 6.40 (br s, 1H), 5.66 (br d, J = 8.7 Hz, 1H), 4.42 - 4.21 (m, 2H), 4.16 (br dd, J = 5.0, 8.3 Hz, 1H), 3.79 (s, 3H), 3.72 - 3.52 (m, 2H), 3.27 - 3.19 (m, 1H), 3.12 (s, 3H), 2.97 - 2.87 (m, 1H), 2.59 (br d, J = 4.5 Hz, 3H), 2.29 - 2.19 (m, 1H), 2.18 - 2.07 (m, 1H), 1.99 (br d, J = 10.8 Hz, 1H), 1.57 - 1.42 (m, 1H), 1.18 - 1.02 (m, 1H), 0.89 (br s, 3H).

EXAMPLE S101.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (101)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1- (2-morpholinoethyl) indol-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₇H₄₃ClN₁₀O₄ theory 726.3, found 727.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 8.70 (s, 1H), 8.04 (s, 1H), 7.64-7.53 (m, 2H), 7.31 (d, J = 8.80 Hz, 1H), 7.13-7.00 (m, 1H), 6.50 (d, J = 8.80 Hz, 1H), 6.45 (s, 1H), 5.70 (d, J = 8.40 Hz, 1H), 4.52-4.45 (m, 2H), 4.19-4.16 (m, 1H), 4.08-3.90 (m, 2H), 3.81 (s, 3H), 3.65-3.45 (m, 6H), 3.40-3.35 (m, 1H), 3.19-3.11 (s, 1H), 3.05-2.95 (m, 1H), 2.71-2.60 (m, 1H), 2.59-2.54 (m, 2H), 2.46-2.42 (m, 1H), 2.31-2.22 (m, 1H), 2.19-2.08 (m, 2H), 1.62-1.55 (m, 1H), 1.21-1.15 (m, 1H), 0.98 (d, J = 6.0 Hz, 3H).

EXAMPLE S102 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (102)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S101 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak. LCMS C ₃₇H₄₃ClN₁₀O₄ theory 726.3, found 727.2 [M+H]⁺;¹H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.64 (s, 1H), 8.02 (s, 1H), 7.56 (br s, 1H), 7.50 (br d, J = 8.3 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.99 (d, J = 8.3 Hz, 1H), 6.50 (dd, J = 1.5, 8.9 Hz, 1H), 6.43 (s, 1H), 5.68 (d, J = 8.9 Hz, 1H), 4.51 - 4.41 (m, 2H), 4.17 (dd, J = 5.2, 8.6 Hz, 1H), 3.81 (s, 3H), 3.76 (br t, J = 6.5 Hz, 2H), 3.56 - 3.48 (m, 5H), 3.02 (br t, J = 12.3 Hz, 1H), 2.73 - 2.57 (m, 4H), 2.42 (br s, 3H), 2.30 - 2.01 (m, 4H), 1.64 - 1.51 (m, 1H), 1.23 - 1.11 (m, 1H), 0.96 (br d, J = 6.4 Hz, 3H).

EXAMPLE S103 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (103)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2-morpholinoethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S101 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the second elution peak. LCMS C ₃₇H₄₃ClN₁₀O₄ theory 726.3, found 727.2 [M+H]⁺;¹H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.65 (s, 1H), 8.02 (s, 1H), 7.56 (s, 1H), 7.50 (br d, J = 8.5 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.99 (br d, J = 8.5 Hz, 1H), 6.50 (dd, J = 1.5, 8.8 Hz, 1H), 6.43 (s, 1H), 5.68 (d, J = 9.1 Hz, 1H), 4.46 (br s, 2H), 4.17 (dd, J = 5.2, 8.8 Hz, 1H), 3.81 (s, 3H), 3.76 (br t, J = 6.2 Hz, 2H), 3.52 (br d, J = 16.2 Hz, 6H), 3.02 (br t, J = 12.6 Hz, 1H), 2.74 - 2.56 (m, 4H), 2.42 (br s, 3H), 2.31 - 2.03 (m, 3H), 1.63 - 1.53 (m, 1H), 1.22 - 1.12 (m, 1H), 0.97 (br d, J = 6.3 Hz, 3H).

EXAMPLE S104.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2- (methylamino) ethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (104)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2- (methylamino) ethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1, 6 and 5 using (2- (5-amino-2-oxoindol-1-yl) ethyl) (methyl) carbamic acid tert-butyl ester and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₄H₃₉ClN₁₀O₃ theory 670.3, found 672.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.84 (s, 1H), 8.67 (s, 1H), 8.03 (s, 1H), 7.59 (s, 1H), 7.51 (d, J = 8.40, 1.60 Hz, 1H), 7.32 (d, J = 8.40 Hz, 1H), 7.01 (d, J = 8.40 Hz, 1H), 6.50 (dd, J = 8.80, 1.60 Hz, 1H), 6.45 (s, 1H), 5.69-5.58 (m, 1H), 4.55-4.47 (m, 2H), 4.19-4.16 (m, 1H), 3.81 (s, 3H), 3.80-3.75 (m, 2H), 3.53 (s, 2H), 3.05-2.96 (s, 1H), 2.85-2.75 (m, 2H), 2.72-2.65 (m, 1H), 2.62-2.55 (m, 2H), 2.36 (s, 3H), 2.28-2.20 (m, 2H), 2.19-2.05 (m, 1H), 1.62-1.53 (m, 1H), 1.25-1.18 (m, 1H), 0.97 (d, J = 6.4 Hz, 3H).

EXAMPLE S105 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2- (methylamino) ethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (105)

The product of example S104 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=18.13 min. LCMS C ₃₄H₃₉ClN₁₀O₃ theory 670.3, found 672.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.80 (s, 1H), 8.64 (s, 1H), 8.02 (s, 1H), 7.56 (s, 1H), 7.50 (dd, J = 1.8, 8.5 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.99 (d, J = 8.5 Hz, 1H), 6.50 (dd, J = 1.7, 8.8 Hz, 1H), 6.44 (s, 1H), 5.68 (d, J = 9.1 Hz, 1H), 4.53 - 4.41 (m, 2H), 4.17 (dd, J = 5.1, 8.8 Hz, 1H), 3.81 (s, 3H), 3.71 (t, J = 6.4 Hz, 2H), 3.52 (s, 2H), 3.07 - 2.96 (m, 1H), 2.73 - 2.57 (m, 7H), 2.32 - 2.20 (m, 5H), 2.19 - 2.02 (m, 3H), 1.64 - 1.54 (m, 1H), 1.20 - 1.11 (m, 1H), 0.97 (d, J = 6.4 Hz, 3H).

EXAMPLE S106 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2- (methylamino) ethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (106)

The product of example S104 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt= 40.27 min. LCMS C ₃₄H₃₉ClN₁₀O₃ theory 670.3, found 672.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.80 (s, 1H), 8.64 (s, 1H), 8.01 (s, 1H), 7.56 (s, 1H), 7.50 (dd, J = 1.7, 8.4 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 6.99 (d, J = 8.6 Hz, 1H), 6.50 (dd, J = 1.5, 8.8 Hz, 1H), 6.44 (s, 1H), 5.68 (d, J = 9.1 Hz, 1H), 4.53 - 4.39 (m, 2H), 4.17 (dd, J = 5.1, 8.7 Hz, 1H), 3.81 (s, 3H), 3.70 (t, J = 6.5 Hz, 2H), 3.52 (s, 2H), 3.02 (br t, J = 11.7 Hz, 1H), 2.74 - 2.58 (m, 7H), 2.31 - 2.20 (m, 4H), 2.19 - 2.03 (m, 2H), 1.58 (br dd, J = 4.5, 5.8 Hz, 1H), 1.22 - 1.12 (m, 1H), 0.97 (d, J = 6.4 Hz, 3H).

EXAMPLE S107.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (107)

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1- (3-fluoro-3-methylbutyl) indol-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₆H₄₁ClFN₉O₃ theory of 701.3, found 702.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.82 (s, 1H), 9.62 (s, 1H), 8.15 (s, 1H), 7.51 (s, 1H), 7.46 (d, J = 8.40 Hz, 1H), 7.33 (d, J = 8.80 Hz, 1H), 6.97 (d, J = 8.40 Hz, 1H), 6.53 (d, J = 8.80 Hz, 1H), 6.47 (s, 1H), 4.27-4.25 (m, 2H), 4.19-4.16 (m, 1H), 3.81 (s, 3H), 3.54 (s, 2H), 3.36-3.31 (m, 1H), 3.18-3.12 (m, 1H), 2.88-2.82 (m, 1H), 2.63-2.58 (m, 2H), 2.26-2.23 (m, 1H), 2.16-2.09 (m, 2H), 1.92-1.83 (m, 2H), 1.65-1.63 (m, 1H), 1.36 (d, J = 21.60 Hz, 6H), 1.24-1.22 (m, 1H), 0.96 (d, J = 6.40 Hz, 3H).

EXAMPLE S108 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (108)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S107 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=7.46 min. LCMS C ₃₆H₄₁ClFN₉O₃ theory of 701.3, found 702.2 [M+H]⁺;¹H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.70 (br s, 1H), 8.03 (s, 1H), 7.59 (s, 1H), 7.51 (dd, J = 1.8, 8.5 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.50 (dd, J = 1.4, 8.8 Hz, 1H), 6.43 (s, 1H), 5.68 (br d, J = 8.9 Hz, 1H), 4.52 - 4.39 (m, 2H), 4.17 (dd, J = 5.1, 8.7 Hz, 1H), 3.81 (s, 3H), 3.78 - 3.72 (m, 2H), 3.54 (s, 2H), 3.02 (br t, J = 12.2 Hz, 1H), 2.73 - 2.56 (m, 4H), 2.31 - 2.04 (m, 4H), 1.96 - 1.84 (m, 2H), 1.64 - 1.52 (m, 1H), 1.40 (s, 3H), 1.35 (s, 3H), 1.26 - 1.10 (m, 2H), 0.96 (d, J = 6.4 Hz, 3H).

EXAMPLE S109 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (109)

Step 1 synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (3-fluoro-3-methylbutyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S107 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22 ℃. The title compound was isolated as a second elution peak, rt=10.56 min. LCMS C ₃₆H₄₁ClFN₉O₃ theory of 701.3, found 702.2 [M+H]⁺;¹H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.68 (br s, 1H), 8.02 (s, 1H), 7.59 (s, 1H), 7.51 (dd, J = 1.8, 8.5 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 6.92 (d, J = 8.5 Hz, 1H), 6.51 (dd, J = 1.4, 8.8 Hz, 1H), 6.43 (s, 1H), 5.68 (br d, J = 9.1 Hz, 1H), 4.53 - 4.39 (m, 2H), 4.17 (dd, J = 5.2, 8.9 Hz, 1H), 3.81 (s, 3H), 3.77 - 3.70 (m, 2H), 3.53 (s, 2H), 3.02 (br t, J = 12.3 Hz, 1H), 2.72 - 2.58 (m, 4H), 2.30 - 2.03 (m, 4H), 1.94 - 1.83 (m, 2H), 1.63 - 1.53 (m, 1H), 1.40 (s, 3H), 1.35 (s, 3H), 1.28 - 1.12 (m, 2H), 0.96 (d, J = 6.4 Hz, 3H).

EXAMPLE S110.3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2, 2-difluoropropyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (110)

The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1- (2, 2-difluoropropyl) indol-2-one and 3- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione as starting materials. LCMS C ₃₄H₃₆ClF₂N₉O₃ theory 691.3, found 692.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.98 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.59 (s, 1H), 7.52 (dd, J = 2.00 Hz and 8.4 Hz, 1H), 7.31 (d, j=8.80 Hz, 1H), 7.03 (d, j=8.80 Hz, 1H), 6.51 (dd, j=1.60 Hz and 8.8 Hz, 1H), 6.45 (s, 1H), 5.70 (d, J = 8.80 Hz, 1H), 4.54-4.46 (m, 2H), 4.19-4.10 (m, 3H), 3.82 (s, 3H), 3.65 (s, 2H), 3.02 (t, J = 12.80 Hz, 1H), 2.69-2.65 (m, 2H), 2.29-2.22 (m, 1H), 2.18-2.12 (m, 1H), 2.09-2.06 (m, 1H), 1.67 (t, J = 19.2 Hz, 3H), 1.62-1.58 (m, 2H), 1.18-1.13 (m, 1H), 0.97 (d, J = 6.80 Hz, 4H).

EXAMPLE S111 Synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2, 2-difluoropropyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (111)

Step 1 synthesis of (R) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2, 2-difluoropropyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The product of example S110 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS, 21X 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additives)) at 22 ℃. The title compound was isolated as the first eluting peak, rt=6.18 min. LCMS C ₃₄H₃₆ClF₂N₉O₃ theory 691.3, found 692.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.80 (s, 1H), 8.68 (s, 1H), 8.02 (s, 1H), 7.58 (s, 1H), 7.51 (dd, J = 1.9, 8.6 Hz, 1H), 7.30 (d, J = 8.7 Hz, 1H), 7.02 (d, J = 8.3 Hz, 1H), 6.50 (dd, J = 1.6, 8.8 Hz, 1H), 6.44 (s, 1H), 5.68 (br d, J = 9.1 Hz, 1H), 4.54 - 4.39 (m, 2H), 4.26 - 4.06 (m, 3H), 3.81 (s, 3H), 3.64 (s, 2H), 3.02 (br t, J = 11.8 Hz, 1H), 2.74 - 2.56 (m, 4H), 2.38 - 2.34 (m, 1H), 2.30 - 2.03 (m, 4H), 1.66 (t, J = 19.1 Hz, 3H), 1.60 - 1.53 (m, 1H), 1.22 - 1.13 (m, 1H), 0.96 (d, J = 6.4 Hz, 3H).

EXAMPLE S112 Synthesis of (S) -3- (6- (((3R, 4R) -1- (5-chloro-4- ((1- (2, 2-difluoropropyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (112)

The product of example S110 was then isolated using preparative HPLC (with REGIS WHELK-O1 SS,21 x 250 mm,5 u column, 1 mL/min flow rate, isocratic elution in 100% MeCN (no additive)) at 22oC the title compound was isolated as a second elution peak, rt=8.48 min. LCMS C ₃₄H₃₆ClF₂N₉O₃ theory 691.3, found 692.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.80 (s, 1H), 8.66 (s, 1H), 8.02 (s, 1H), 7.58 (s, 1H), 7.54 - 7.48 (m, 1H), 7.30 (d, J = 8.7 Hz, 1H), 7.02 (d, J = 8.7 Hz, 1H), 6.53 - 6.48 (m, 1H), 6.44 (s, 1H), 5.68 (d, J = 9.1 Hz, 1H), 4.52 - 4.40 (m, 2H), 4.20 - 4.08 (m, 3H), 3.81 (s, 3H), 3.64 (s, 2H), 3.07 - 2.96 (m, 1H), 2.73 - 2.58 (m, 6H), 2.38 - 2.34 (m, 1H), 2.31 - 2.03 (m, 3H), 1.66 (t, J = 19.1 Hz, 3H), 1.60 - 1.53 (m, 1H), 1.23 - 1.12 (m, 1H), 0.96 (d, J = 6.4 Hz, 3H).

The examples in table 3 below were prepared according to the general procedure outlined in the table starting with their respective commercial starting materials and intermediates found in this document.

EXAMPLE S124.Synthesis of 3- [6- [1- [ 5-chloro-4- [ (2-oxoindol-5-yl) amino ] pyrimidin-2-yl ] -4-piperidinyl ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione (124)

Step 1 Synthesis of 4- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] piperidine-1-carboxylic acid tert-butyl ester. 3- [ 6-benzyloxy-2- (phenoxymethyl) -3-pyridinyl ] -6-bromo-1-methyl-indazole (300 mg,0.60 mmol), 4-bromopiperidine-1-carboxylic acid tert-butyl ester (250 mg,0.95 mmol), [4,4' -bis (1, 1-dimethylethyl) -2,2' -bipyridyl-N1, N1' ] bis [3, 5-difluoro-2- [5- (trifluoromethyl) -2-pyridinyl-N ] phenyl-C ] iridium (III) hexafluorophosphate (6.73 mg,0.0100 mmol), tris (trimethylsilyl) silane (149.08 mg,0.6000 mmol) and lithium hydroxide (28.72 mg,1.2 mmol) were added to a stirring-rod-equipped vial. Sealed and purged with nitrogen, then 1, 4-dioxane (2 mL) was added. The pre-catalyst solution was prepared separately. Nickel (II) chloride ethylene glycol dimethyl ether complex (6.59 mg,0.030 mmol) and 4,4 '-di-tert-butyl-2, 2' -bipyridine (8.05 mg,0.030 mmol) were added to a vial equipped with a stirring rod. Sealed and purged with nitrogen. 1, 4-dioxane (2 mL) was added and 10 was stirred/sonicated min. 0.2 mL precatalyst solution was added to the initial reaction vial. Thereafter, purged with argon and stirred for 10 minutes. Sealed with parafilm and irradiated overnight at room temperature with a 34W blue LED lamp. LCMS indicated completion of the reaction. The mixture was concentrated and purified by silica gel chromatography using 0-40% EA/hexane to give the title compound as a pale yellow oil (208 mg,0.336 mmol,56.0% yield).

Step 2 Synthesis of 3- [6- [1- [ 5-chloro-4- [ (2-oxoindol-5-yl) amino ] pyrimidin-2-yl ] -4-piperidinyl ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione the title compound was synthesized according to general procedures 4, 5 and 6 using 4- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₀H₂₉ClN₈O₃ theory 584.2, found 585.3 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 10.30 (s, 1H), 8.59 (s, 1H), 8.01 (s, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.48 (s, 1H), 7.43 (s, 1H), 7.40 (dd, J = 8.4, 2.0 Hz, 1H), 7.04 (d, J = 7.9 Hz, 1H), 6.76 (d, J = 8.3 Hz, 1H), 4.72 – 4.62 (m, 2H), 4.32 (dd, J = 9.7, 5.1 Hz, 1H), 3.95 (s, 3H), 3.46 (s, 2H), 2.97 – 2.87 (m, 3H), 2.69 – 2.55 (m, 2H), 2.38 – 2.29 (m, 1H), 2.21 – 2.12 (m, 1H), 1.86 (d, J = 11.9 Hz, 2H), 1.70 – 1.58 (m, 2H).

EXAMPLE S125.3 Synthesis of 3- (6- (1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (125)

Step 1 Synthesis of 3- (6- (1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -piperidin-4-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione the title compound was synthesized according to general procedures 4, 5 and 6 using 4- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₀H₂₉ClN₈O₃ theory 598.2, found 599.3 [M+H]⁺;¹H NMR (DMSO-d₆, 400 MHz) δ 10.8-10.9 (m, 1H), 8.6-8.7 (m, 1H), 8.03 (s, 1H), 7.6-7.6 (m, 1H), 7.5-7.6 (m, 2H), 7.4-7.5 (m, 1H), 7.0-7.1 (m, 1H), 6.9-7.0 (m, 1H), 4.6-4.7 (m, 2H), 4.3-4.4 (m, 1H), 3.9-4.0 (m, 3H), 3.5-3.6 (m, 2H), 3.1-3.1 (m, 3H), 2.9-3.0 (m, 3H), 2.5-2.7 (m, 2H), 2.3-2.4 (m, 1H), 2.1-2.2 (m, 1H), 1.8-1.9 (m, 2H), 1.6-1.7 (m, 2H).

EXAMPLE S126.Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (126)

Step 1 Synthesis of tert-butyl N- [1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -2-oxo-4-piperidinyl ] carbamate. A mixture of tert-butyl N- (2-oxo-4-piperidinyl) carbamate (1.3 g,6.06 mmol), 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole （3.33 g,6.66 mmol）、Pd₂(dba)₃（277 mg,0.30 mmol）、Me₄tBuXphos（291 mg,0.61 mmol） and K ₃PO₄ (3.21 g,15.2 mmol) in tBuOH (60 mL) was heated to 100℃for 24 h and then cooled to room temperature. The mixture was filtered over celite and washed with DCM (15 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-10% MeOH in DCM. The fractions were concentrated and the residue was further purified by reverse phase chromatography (C18) using a gradient of 0-100% MeCN and 10 mM ammonium formate in water to give the title compound as a solid （2.64 g,69%）.MS (ESI) [M+H]⁺ 634.7.¹H NMR (500 MHz, DMSO-d₆) δ 7.91 (d, J = 8.1 Hz, 1H), 7.66 (d, J = 8.7 Hz, 1H), 7.56 (d, J = 1.1 Hz, 1H), 7.50 – 7.45 (m, 2H), 7.44 – 7.25 (m, 8H), 7.18 (s, 1H), 6.97 (dd, J = 8.7, 1.7 Hz, 1H), 6.60 (d, J = 8.1 Hz, 1H), 5.48 (s, 2H), 5.43 (s, 2H), 4.05 (s, 3H), 3.91 (s, 1H), 3.77 – 3.57 (m, 2H), 2.78 – 2.60 (m, 1H), 2.40 – 2.30 (m, 1H), 2.09 (d, J = 6.0 Hz, 1H), 1.93 – 1.75 (m, 1H), 1.43 (s, 9H).

Step 2 Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was accomplished using general procedures 4, 5 and 6 using N- [1- [3- (2, 6-dibenzyloxy-3-pyridyl) -1-methyl-indazol-6-yl ] -2-oxo-4-piperidinyl ] carbamic acid tert-butyl ester as starting material. The crude residue was purified by reverse phase chromatography (C18) using a gradient of 0-100% MeCN and 10mM ammonium formate in water. The fractions were concentrated under reduced pressure and the residue was further purified by preparative HPLC (BEH column, C18) using a gradient of 25% -35% MeCN and 10 mM% ammonium formate in water to give the title compound (8.6 mg, 5%) as a solid. LCMS C ₃₁H₃₀ClN₉O₄ theory 627.2, found 628.2 [M+H]⁺.¹H NMR (500 MHz, DMSO-d₆) δ 10.91 (s, 1H), 8.60 (s, 1H), 7.98 (s, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.55 (s, 1H), 7.51 (d, J = 8.1 Hz, 1H), 7.20 (s, 1H), 7.03 (d, J = 8.7 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 4.38 (dd, J = 10.1, 5.0 Hz, 1H), 3.98 (s, 3H), 3.69 (s, 2H), 3.57 (s, 2H), 3.12 (s, 3H), 2.78 – 2.58 (m, 4H), 2.47 – 2.33 (m, 2H), 2.23 – 2.15 (m, 2H), 1.96 – 1.88 (m, 1H).

EXAMPLE S127.Synthesis of 3- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (127)

Step 1 Synthesis of 4- [ benzyl (methyl) amino ] piperidin-2-one to a solution of piperidine-2, 4-dione (1.0 g,8.8 mmol) in DCE (25 mL) were added N-methyl-1-phenyl-methylamine (1.1 mL,8.8 mmol) and AcOH (0.5 mL,8.8 mmol) in sequence and the mixture was stirred at room temperature 2 h. At this point NaBH ₃ CN (833 mg,13.3 mmol) was added to the reaction mixture and stirring was continued 36 h. Saturated aqueous NaHCO ₃ (100 mL) and DCM (100 mL) were added and the layers were separated. The organic layer was washed with water (100 mL), brine (20 mL), dried (Na ₂SO₄), filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound as a semi-solid （790 mg,41%）.MS (ESI) [M+H]⁺ 219.2.¹H NMR (400 MHz, DMSO-d₆) δ 7.45 (br s, 1H), 7.35 – 7.28 (m, 4H), 7.27 – 7.19 (m, 1H), 3.54 (q, J = 13.5 Hz, 2H), 3.22 – 3.16 (m, 1H), 3.08 – 2.99 (m, 1H), 2.89 – 2.80 (m, 1H), 2.31 – 2.20 (m, 2H), 2.09 (s, 3H), 1.97 – 1.87 (m, 1H), 1.68 – 1.54 (m, 1H).

Step 2 Synthesis of 4- [ benzyl (methyl) amino ] -1- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] piperidin-2-one to a solution of 4- [ benzyl (methyl) amino ] piperidin-2-one (300 mg,1.24 mmol), 6-bromo-3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazole (681 mg,1.36 mmol, intermediate provided by Celgene), pd ₂(dba)₃ (226 mg,0.25 mmol) and XantPhos (358 mg,0.62 mmol) in 1, 4-dioxane (6.2 mL) was added Cs ₂CO₃ (564 mg,1.73 mmol) at room temperature. The reaction mixture was heated to 100 ℃ for 18 h and then cooled to room temperature. The mixture was diluted with DCM (100 mL), filtered through a pad of celite, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography using a gradient of 0-20% MeOH in DCM to give the title compound as a solid (300 mg, 38%). MS (ESI) [ M+H ] ⁺ 639.3.

Step 3 Synthesis of 3- [6- [4- [ [ 5-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidin-2-yl ] -methyl-amino ] -2-oxo-1-piperidinyl ] -1-methyl-indazol-3-yl ] piperidine-2, 6-dione the synthesis of the title compound was completed using general procedures 4 and 6 using 4- [ benzyl (methyl) amino ] -1- [3- (2, 6-dibenzyloxy-3-pyridinyl) -1-methyl-indazol-6-yl ] piperidin-2-one as starting material. The final fraction was heated to 100 ℃ for 18 h. Purification was then performed by preparative HPLC (BEH column, C18) using a gradient of 26% -36% MeCN and 10 mM ammonium formate in water to give the title compound (13.6 mg, 12%) as a solid. LCMS C ₃₂H₃₂ClN₉O₄ theory 641.1, found 642.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.90 (br s, 1H), 8.68 (s, 1H), 8.04 (s, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.63 (s, 1H), 7.56 – 7.47 (m, 2H), 6.98 (d, J = 8.2 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 4.38 (dd, J = 10.1, 5.0 Hz, 1H), 3.98 (s, 3H), 3.83 – 3.72 (m, 1H), 3.71 – 3.59 (m, 1H), 3.54 (d, J = 4.9 Hz, 2H), 3.29 (s, 1H), 3.09 (s, 3H), 3.00 (s, 3H), 2.76 – 2.60 (m, 4H), 2.44 – 2.31 (m, 1H), 2.28 – 2.14 (m, 2H), 2.06 – 1.96 (m, 1H).

EXAMPLE S128.3 Synthesis of 3- (6- ((S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (128)

Step 1 Synthesis of 3- (6- ((S) -4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized using chiral SFC using the product of example S128 and using the first eluting isomer in general procedures 4,5 and 6. Rt 6.63 min, sfc column: lux C4, mobile phase: 55:45 (a: B), a=liquid CO ₂, b=0.5% isopropylamine in methanol, flow rate ：5.0 mL/min;MS (ESI) 642.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6): δ 10.93 (s, 1H), 8.71 (s, 1H), 8.06 (s, 1H), 7.70 (d, J = 8.40 Hz, 1H), 7.64 (s, 1H), 7.54-7.52 (m, 2H), 6.99-6.94 (m, 2H), 4.94 (brs, 1H), 4.41-4.37 (m, 1H), 3.99 (s, 3H), 3.82-3.75 (m, 1H), 3.67-3.62 (m, 1H), 3.55-3.54 (m, 2H), 3.10 (s, 3H), 3.00 (s, 3H), 2.68-2.59 (m, 2H), 2.46-2.38 (m, 1H), 2.25-2.15 (m, 2H), 2.05-1.95 (m, 1H).

Example S129.3- (6- ((R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (129) Synthesis

Step 1 Synthesis of 3- (6- ((R) -4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized using chiral SFC using the product of example S128 and using the second eluting isomer of general procedures 4,5 and 6. Rt 7.46 min, sfc column: lux C4, mobile phase: 55:45 (a: B), a=liquid CO ₂, b=0.5% isopropylamine in methanol, flow rate ：5.0 mL/min.MS (ESI) 642.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d6): δ 10.92 (s, 1H), 8.70 (s, 1H), 8.05 (s, 1H), 7.70 (d, J = 8.40 Hz, 1H), 7.64 (s, 1H), 7.53-7.51 (m, 2H), 6.99-6.94 (m, 2H), 4.95 (brs, 1H), 4.41-4.37 (m, 1H), 3.99 (s, 3H), 3.81-3.76 (m, 1H), 3.67-3.64 (m, 1H), 3.55-3.54 (m, 2H), 3.10 (s, 3H), 3.00 (s, 3H), 2.72-2.64 (m, 2H), 2.46-2.39 (m, 1H), 2.23-2.18 (m, 2H), 2.03-1.98 (s, 1H).

EXAMPLE S130.Synthesis of 3- (6- ((3R, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -3-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (130)

Step 1 (S) -Synthesis of 4- (benzyl (methyl) amino) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) piperidin-2-one (isomer 2) A stirred solution of 4- [ benzyl (methyl) amino ] piperidin-2-one (3.66 g,16.8 mmol) and 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazol (7.0 g,14.0 mmol) in 1, 4-dioxane (150 mL) was degassed with nitrogen gas 5 min. Pd ₂(dba)₃ (1.28 g,1.40 mmol), xantphos (1.619 g,2.80 mmol) and Cs ₂CO₃ (6.84 g,20.98 mmol) were then added. The reaction mixture was heated to 100 ℃ and stirred 16 h. The reaction mixture was treated with water (200 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel with 10% methanol in dichloromethane to give the racemate as an off-white solid (3.50 g). The racemate was separated by chiral preparative HPLC to give isomer 1 (Rt 8.73 min) and the title compound as an off-white solid as the second eluting isomer (isomer-2,1.55 g,2.408 mmol,17% yield). Chiral HPLC method, column CHIRALPAK IJ (250X 4.6 mm), 5.0 μm, 0.2% ammonia in methanol/isopropanol, flow rate 3.0 mL/min, rt 9.87 min. MS (ESI) [ M+H ] ⁺ 638.3.

Step 2 Synthesis of (3R, 4S) -4- (benzyl (methyl) amino) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3-methylpiperidin-2-one. To a stirred solution of the compound (S) -4- (benzyl (methyl) amino) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) piperidin-2-one (250 mg,0.392 mmol) in THF (8.0 mL) under nitrogen was added dropwise 1.0M LHMDS (0.784 mL,0.784 mmol) in hexane and stirred 45 min. Methyl iodide (0.049 mL,0.784 mmol) was then added and warmed slowly to 25 ℃ with stirring 1 h. The reaction mixture was treated with saturated NH ₄ Cl solution (30 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product. The crude product was purified by flash column chromatography on silica gel with 50% -70% ethyl acetate/petroleum ether to give the title compound as an off-white solid (180 mg,0.271 mmol,69% yield ）.MS (ESI) [M+H]⁺ 652.3; ¹H NMR (400 MHz, DMSO-d₆): δ 7.91 (d, J = 8.8 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 7.53 (s, 1H), 7.49-7.46 (m, 2H), 7.41-7.24 (m, 12 H), 6.93 (dd, J = 1.6 Hz and 8.8 Hz, 1H), 6.59 (d, J = 8.0 Hz, 1H), 5.47 (s, 2H), 5.42 (s, 2H), 4.04 (s, 3H), 3.78 (d, J = 13.6 Hz, 1H), 3.68-3.66 (m, 2H), 3.52 (d, J = 13.6 Hz, 1H), 2.80-2.76 (m, 1H), 2.68-2.63 (m, 1H), 2.19 (s, 3H), 2.18-2.13 (m, 1H), 1.96-1.91 (m, 1H), 1.28 (d, J = 6.8 Hz, 1H).

Step 3 Synthesis of 3- (1-methyl-6- ((3R, 4S) -3-methyl-4- (methylamino) -2-oxopiperidin-1-yl) -1H-indazol-3-yl) piperidine-2, 6-dione. To a stirred solution of (3 r,4 s) -4- (benzyl (methyl) amino) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -3-methylpiperidin-2-one (120 mg,0.180 mmol) in ethanol (2.5 mL) and THF (2.5 mL) under nitrogen was added TFA (0.014 L,0.180 mmol) and 20% palladium on carbon hydroxide (60 mg). The reaction mixture was stirred at 50 ℃ under hydrogenation pressure (5 atm) for 16: 16 h. The reaction mixture was filtered through a pad of celite and the pad was washed with a 1:1 mixture of THF and ethanol (30 mL). The filtrate was concentrated under reduced pressure to give the title compound as crude product (70 mg,0.172 mmol,96% yield). The crude product was used in the next step without further purification. MS (ESI) 384.2 [ M+H ] ⁺.

Step 4 Synthesis of 3- (6- ((3R, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -3-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. To a stirred solution of compound 3- (1-methyl-6- ((3R, 4S) -3-methyl-4- (methylamino) -2-oxopiperidin-1-yl) -1H-indazol-3-yl) piperidine-2, 6-dione (50 mg,0.121 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (36.2 mg,0.121 mmol) in DMSO (1.0 mL) was added DIPEA (0.064 mL,0.364 mmol) at 25 ℃. The reaction mixture was heated to 80 ℃ and stirred 5 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC to give the title compound as an off-white solid (8 mg,0.012 mmol,10% yield). LCMS C ₃₃H₃₄ClN₉O₄ theory 655.2, found 656.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.93 (s, 1H), 8.93 (brs, 1H), 8.07 (s, 1H), 7.69 (d, J = 8.8 Hz, 1H), 7.63-7.48 (m, 3H), 6.99-6.96 (m, 2H), 4.81 (brs, 1H), 4.41-4.37 (m, 1H), 3.99 (s, 3H), 3.69-3.62 (m, 2H), 3.54 (s, 2H), 3.11 (s, 3H), 2.97 (s, 3H), 2.21 - 2.18 (m, 2H), 1.99-1.90 (m, 1H), 1.02 (d, J = 6.8 Hz, 3H).

EXAMPLE S131.Synthesis of 3- (6- ((3S, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -3-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (131)

Step 1 Synthesis of 3- (6- ((3S, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -3-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was accomplished by the procedure outlined for example S131 and using the first eluting isomer from example S131 step 1. LCMS C ₃₃H₃₄ClN₉O₄ theory 655.2, found 656.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 10.62 - 10.53 (m, 1H), 8.42 (s, 1H), 8.03 (s, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.61 (s, 1H), 7.52 (br d, J = 8.8 Hz, 1H), 7.45 (s, 1H), 6.97 (dd, J = 1.4, 8.6 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 4.87 - 4.77 (m, 1H), 4.36 (dd, J = 5.3, 9.3 Hz, 1H), 3.99 (s, 3H), 3.76 - 3.66 (m, 2H), 3.53 (s, 2H), 3.16 - 3.12 (m, 3H), 3.01 - 3.00 (m, 3H), 2.71 - 2.66 (m, 2H), 2.44 - 2.33 (m, 2H), 2.29 - 2.19 (m, 2H), 2.05 - 1.96 (m, 1H), 1.07 (d, J = 7.0 Hz, 3H).

EXAMPLE S132.2 Synthesis of- ((3R, 4R) -4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidin-1-yl) -4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidine-5-carbonitrile (132)

Step 1 Synthesis of 1- (1-methyl-6-nitro-indazol-3-yl) hexahydropyrimidine-2, 4-dione. A mixture of 3-bromo-1-methyl-6-nitro-indazole (3.00 g,11.7 mmol), hexahydropyrimidine-2, 4-dione (1.60 g,14.1 mmol), tripotassium phosphate (6.22 g,29.3 mmol), tetramethyl tBuXPhos (281 mg,0.58 mmol) and Pd ₂(dba)₃ (268 mg,0.29 mmol) in t-butanol (125 mL) was degassed 3 times with nitrogen. The reaction mixture was heated to 100 ℃ with stirring for 24 h and then cooled to room temperature. Additional Pd ₂(dba)₃ (268 mg,0.29 mmol) and tetramethyl tBuXPhos (281 mg,0.58 mmol) were added. The reaction mixture was heated to 100 ℃ for a further 16 h and then cooled to room temperature. The volatiles were evaporated under reduced pressure. Water (200 mL) was added and the resulting precipitate was collected by filtration on a Buchner funnel. The solid was then washed with water (3×50 mL) and dried under vacuum. The material was triturated in diethyl ether (100 mL) and collected by filtration to give the title compound (2.64 g, 78%) as a solid which was used in the next step without further purification. LCMS C ₁₂H₁₁N₅O₄ theory 289.1, found 290.2 [M+H] +;¹H NMR (500 MHz, DMSO) δ 10.67 (s, 1H), 8.70 (dd, J = 1.7, 0.7 Hz, 1H), 7.95 – 7.84 (m, 2H), 4.14 (s, 3H), 3.97 (t, J = 6.7 Hz, 2H), 2.77 (t, J = 6.7 Hz, 2H).

Step 2 Synthesis of 1- (6-amino-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione. A mixture of 1- (1-methyl-6-nitro-indazol-3-yl) hexahydropyrimidine-2, 4-dione (2.64 g,9.13 mmol) and 10% Pd/C (0.97 g,0.91 mmol) in methanol (250 mL) was hydrogenated under a hydrogen atmosphere (1 atm) at room temperature for 6.5 h. The mixture was filtered through celite and washed with MeOH (5 x 45 mL). The filtrate was concentrated under reduced pressure, the material was triturated in ether and collected by filtration to give the title (1.92 g, 81%) as a solid.

Step 3 Synthesis of 1- (6-iodo-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione. To a solution of 1- (6-amino-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione (2.1 g,8.1 mmol) in acetic acid (21 mL) cooled to 0 ℃ was added a solution of H ₂SO₄ (1.11 mL,20.3 mmol) in water (5 mL) followed by a solution of sodium nitrite (838 mg,12.15 mmol) in water (5 mL). The reaction mixture was stirred at 0 ℃ for 2 h. At this point, a solution of KI (4.0 g,24 mmol) in water (5 mL) was added and the mixture was stirred at 0 ℃ for 2: 2 h, then warmed to room temperature. A 50% aqueous solution of NaHSO ₃ (50 mL) was added to the mixture and 18 h was stirred. The resulting precipitate was collected by filtration, washed with water (3×10 mL) and Et ₂ O (3×10 mL), then dried under vacuum to give the title compound (1.98 g, 66%) as a solid, which was used in the next step without further purification. Theoretical value of LCMS C ₁₂H₁₁IN₄O₂ is 370.0, found value is 371.3 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.58 (s, 1H), 8.12 (d, J = 0.6 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 7.39 (dd, J = 8.6, 1.3 Hz, 1H), 3.97 (s, 3H), 3.92 (t, J = 6.7 Hz, 2H), 2.75 (t, J = 6.7 Hz, 2H).

Step 4 Synthesis of 1- [ 1-methyl-6- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) indazol-3-yl ] hexahydropyrimidine-2, 4-dione. A mixture of 1- (6-iodo-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione (500 mg,1.35 mmol), B ₂pin₂（412 mg,1.62 mmol）、Pd(OAc)₂ (30 mg,0.14 mmol) and KOAc (398 mg,4.05 mmol) in DMF (14 mL) was heated to 80 ℃ for 18 h and then cooled to room temperature. The mixture was filtered through celite and washed with EtOAc (5×15 mL). Water (50 mL) was added to the filtrate and the layers were separated. The organic layer was washed with water (5×25 mL), brine (25 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 50% -100% EtOAc in hexanes to give the title compound (335 mg, 67%) as a solid. LCMS C ₁₈H₂₃BN₄O₄ theory was 372.2 and found to be 371.2 m+h ⁺.

Step 5 Synthesis of [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] boronic acid. To a solution of 1- [ 1-methyl-6- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) indazol-3-yl ] hexahydropyrimidine-2, 4-dione (403 mg,1.09 mmol) in THF (5 mL) and water (5 mL) was added NaIO ₄ (698 mg,3.27 mmol) and 1M HCl in water (2.2 ml,2.2 mmol) in sequence. The reaction mixture was stirred at room temperature 18 h. The volatiles were evaporated under reduced pressure and the resulting precipitate was collected by filtration, washed with water (3 x 2 mL) and Et ₂ O (3 x 1 mL), then dried under vacuum to give the title compound as a solid (240 mg, 76%) which was used in the next step without further purification. LCMS C ₁₂H₁₃BN₄O₄ theory 288.1, found 289.1[M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.54 (s, 1H), 8.16 (s, 2H), 8.01 (s, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.52 (d, J = 8.3 Hz, 1H), 4.00 (s, 3H), 3.92 (t, J = 6.7 Hz, 2H), 2.76 (t, J = 6.6 Hz, 2H).

Step 6 Synthesis of tert-butyl (3R, 4R) -4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylate. A mixture of [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] boronic acid (200 mg,0.69 mmol), (3R, 4R) -4-amino-3-methyl-piperidine-1-carboxylic acid tert-butyl ester (228 mg,1.04 mmol), cu (OAc) ₂（152 mg,0.76 mmol）、Et₃ N (190. Mu.L, 1.39 mmol) and 3-A MS (100 mg) in DCE (12.6 mL) was heated to 50℃under O ₂ (1 atm) for 18 h. The mixture was filtered through celite and washed with a 1:1 mixture of MeCN and MeOH (3×10 mL). The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC (BEH column, C18) using a gradient of 10% -80% MeCN and 10mM ammonium formate in water to give the title compound (76 mg, 24%) as a solid. LCMS C ₂₃H₃₂N₆O₄ theory was 456.3 and found to be 457.3 m+h ⁺.

Step 7 Synthesis of 2-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidine-5-carbonitrile.

The title compound was synthesized according to general procedure 1 using 2, 4-dichloropyrimidine-5-carbonitrile as starting material.

Step 8 Synthesis of 2- ((3R, 4R) -4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile. The synthesis of the title compound was completed using general procedures 4 and 5 using (3 r,4 r) -4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] -3-methyl-piperidine-1-carboxylic acid tert-butyl ester and 2-chloro-4- [ (1-methyl-2-oxo-indolin-5-yl) amino ] pyrimidine-5-carbonitrile as starting materials. The crude residue was purified by preparative HPLC (BEH column, C18) using a gradient of 25% -55% MeCN and 10 mM% sodium bicarbonate in water to give the title compound as a solid (6.7 mg, 7%). LCMS C ₃₂H₃₃N₁₁O₃ theory was 619.3, found to be 620.7 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 10.45 (s, 1H), 9.28 (s, 1H), 8.40 (s, 1H), 7.54 – 7.40 (m, 2H), 7.26 (d, J = 8.8 Hz, 1H), 6.94 (d, J = 4.6 Hz, 1H), 6.51 (dd, J = 8.8, 1.9 Hz, 1H), 6.43 (s, 1H), 5.75 (d, J = 8.9 Hz, 1H), 4.66 (br s, 1H), 4.44 (s, 1H), 3.85 (t, J = 6.7 Hz, 2H), 3.81 (s, 3H), 3.53 (s, 2H), 3.18 – 3.03 (m, 3H), 2.85 – 2.75 (m, 1H), 2.72 (t, J = 6.7 Hz, 2H), 2.14 – 2.06 (m, 1H), 1.58 (s, 1H), 1.23 (s, 3H), 0.97 (d, J = 6.4 Hz, 3H).

EXAMPLE S133.1 Synthesis of 1- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) amino) piperidin-1-yl) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (133)

Step 1 Synthesis of 1- (6- (4- ((5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) amino) piperidin-1-yl) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized in analogy to example S133, using tert-butyl N- (4-piperidinyl) carbamate as reactant in step 6. LCMS C ₃₂H₃₃N₁₁O₃ theory 614.2, found 615.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.50 (br s, 1H), 8.53 (br s, 1H), 7.94 (s, 1H), 7.74 (br s, 1H), 7.56 (s, 1H), 7.44 (d, J = 9.0 Hz, 1H), 7.00 (br s, 1H), 6.92 (d, J = 8.8 Hz, 2H), 6.83 (s, 1H), 3.92 – 3.88 (m, 2H), 3.88 (s, 3H), 3.82 (d, J = 12.7 Hz, 2H), 3.69 (br s, 1H), 3.55 (s, 2H), 3.10 (s, 3H), 2.82 (t, J = 11.8 Hz, 2H), 2.73 (t, J = 6.7 Hz, 2H), 1.95 (br d, J = 10.5 Hz, 2H), 1.59 (br dd, J = 21.8, 10.6 Hz, 2H).

EXAMPLE S134.Synthesis of 1- (6- (1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (134)

Step 1 Synthesis of tert-butyl N- (6-bromo-1-methyl-indazol-3-yl) carbamate. To a solution of 6-bromo-1-methyl-indazol-3-amine (1.0 g,4.42 mmol) in 1, 4-dioxane (25 mL) was added Boc ₂ O (1.16 g,5.31 mmol), and the reaction mixture was heated to 100 ℃ for 16 h and then cooled to room temperature. The volatiles were evaporated under reduced pressure and the residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound as a solid （1.18 g,82%）.MS (ESI) [M–tBu]⁺ 271.0.¹H NMR (400 MHz, DMSO-d₆) δ 9.62 (s, 1H), 7.89 (s, 1H), 7.67 (dd, J = 8.6, 2.4 Hz, 1H), 7.20 (dd, J = 8.7, 1.6 Hz, 1H), 3.91 (s, 3H), 1.47 (s, 9H).

Step 2 Synthesis of tert-butyl N- (6-bromo-1-methyl-indazol-3-yl) -N- (2-cyanoethyl) carbamate. To a solution of tert-butyl N- (6-bromo-1-methyl-indazol-3-yl) carbamate (1.18 g,3.61 mmol) in MeCN (15 mL) were added KF (429.4 mg,7.39 mmol), acrylonitrile (0.83 mL,12.6 mmol) and Al ₂O₃ (1.29 g,12.6 mmol) in sequence, and the reaction mixture was heated to reflux for 18 h and then cooled to room temperature. The mixture was filtered over celite and washed with MeCN (20 mL). The filtrate was concentrated under reduced pressure to give the title compound (1.0 g, 73%) as an oil, which was used in the next step without further purification. MS (ESI) [ M-tBu ] ⁺ 324.6.6.

Step 3:N Synthesis of tert-butyl- (3-amino-3-oxo-propyl) -N- (6-bromo-1-methyl-indazol-3-yl) carbamate. To a solution of tert-butyl N- (6-bromo-1-methyl-indazol-3-yl) -N- (2-cyanoethyl) carbamate (12 g,31.6 mmol) in MeOH (90 mL) was added sequentially 30% H ₂O₂ (128 mL,1.23 mol) and NH ₄ OH (118 mL,0.92 mol) at0℃and the reaction mixture was warmed to room temperature and stirred for 6H. Water (500 mL) and DCM (500 mL) were added and the layers separated. The organic layer was washed with Na ₂SO₃ saturated (200 mL) aqueous solution, dried (Na ₂SO₄), filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes, followed by a second purification using and 0-20% MeOH in DCM to give the title compound (10.6 g, 84%) as a solid. MS (ESI) [ M+H ] ⁺ 397.1.

Step 4 Synthesis of 1- (6-bromo-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione. To a solution of tert-butyl N- (3-amino-3-oxo-propyl) -N- (6-bromo-1-methyl-indazol-3-yl) carbamate (10.6 g,26.7 mmol) in THF (125 mL) was added KOTBu (6.0 g,53.4 mmol) and the reaction mixture was stirred at 0℃for 2 h. EtOAc (100 mL) and 1N HCl (40 mL) were added and the layers separated. The organic layer was washed with brine (40 mL), dried (Na ₂SO₄), filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a gradient of 0-100% EtOAc in hexanes to give the title compound (2.1 g, 24%) as a solid. MS (ESI) [ M+H ] ⁺ 323.1.1.

Step 5 Synthesis of 4- [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester. A mixture of 1- (6-bromo-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione (600 mg,1.86 mmol), 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester (603 mg,1.95 mmol), pd (dppf) Cl ₂ ∙ DCM (75.8 mg,90 μmol) and K ₃PO₄ (1.18 g,5.57 mmol) in 1, 4-dioxane (16 mL) and water (4 mL) was heated to 90℃for 4H and then cooled to room temperature. Volatiles were removed and the residue was purified by reverse phase chromatography (C18) with a gradient of 20% -100% MeCN and 10 mM ammonium formate in water to give the title compound as a solid （200 mg,25%）.MS (ESI) [M–H]^–424.3.¹H NMR (500 MHz, DMSO-d₆) δ 10.54 (s, 1H), 7.62 – 7.57 (m, 2H), 7.26 (dd, J = 8.6, 1.4 Hz, 1H), 6.29 (s, 1H), 4.07 – 4.02 (m, 2H), 3.99 (s, 3H), 3.92 (t, J = 6.7 Hz, 2H), 3.61 – 3.55 (m, 2H), 2.76 (t, J = 6.7 Hz, 2H), 2.61 – 2.55 (m, 2H), 1.44 (s, 9H).

Step 6 Synthesis of 1- (6- (1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedures 4, 5 and 6 using 4- [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₀H₃₀ClN₉O₃ theory 599.2, found 600.3 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 10.52 (s, 1H), 8.64 (s, 1H), 8.02 (d, J = 3.7 Hz, 1H), 7.56 – 7.51 (m, 3H), 7.44 (s, 1H), 7.04 (dd, J = 8.6, 1.2 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 4.72 – 4.62 (m, 2H), 3.95 (s, 3H), 3.90 (t, J = 6.7 Hz, 2H), 3.53 (s, 2H), 3.10 (s, 3H), 2.98 – 2.88 (m, 3H), 2.74 (t, J = 6.7 Hz, 2H), 1.90 – 1.83 (m, 2H), 1.71 – 1.58 (m, 2H).

EXAMPLE S135.1 Synthesis of 1- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (135)

Step 1 Synthesis of tert-butyl 4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylate. To a stirred solution of 1- (6-amino-1-methyl-indazol-3-yl) hexahydropyrimidine-2, 4-dione (200 mg,0.77 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (307 mg,1.54 mmol) in DCM (5 mL) was added acetic acid (10 mg,0.77 mmol), the pH was adjusted to 6, and sodium triacetoxyborohydride (490 mg,2.31 mmol) was added portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under nitrogen atmosphere 2 h. The reaction was monitored by LCMS. The reaction mixture was purified rapidly to give the title compound (240 mg, 70%) as a yellow solid.

Step 2 Synthesis of 1- (6- ((1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione the title compound was synthesized according to general procedures 4,5 and 6 using 4- [ [3- (2, 4-dioxohexahydropyrimidin-1-yl) -1-methyl-indazol-6-yl ] amino ] piperidine-1-carboxylic acid tert-butyl ester as starting material. LCMS C ₃₀H₃₁ClN₁₀O₃ theory 614, found 615 [M+H]⁺;¹H NMR (300 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.01 (s, 1H), 7.57 – 7.47 (m, 2H), 7.27 (d, J = 8.8 Hz, 1H), 6.93 (d, J = 9.1 Hz, 1H), 6.55 – 6.44 (m, 2H), 4.38 (d, J = 13.1 Hz, 2H), 3.83 (d, J = 11.9 Hz, 5H), 3.62 – 3.53 (m, 3H), 3.16 – 3.12 (m, 5H), 2.73 – 2.70 (m, 3H), 2.09 – 1.94 (m, 2H), 1.36 – 1.20 (m, 2H).

EXAMPLE S136.Synthesis of 1- (6- ((1- (5-chloro-4- ((2-oxo-1- (2, 2-trifluoroethyl) indol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (136)

Step 1 Synthesis of 1- (6- ((1- (5-chloro-4- ((2-oxo-1- (2, 2-trifluoroethyl) indol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1- (2, 2-trifluoroethyl) indol-2-one and 1- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] hexahydropyrimidine-2, 4-dione as starting materials. LCMS C ₃₁H₃₀ClF₃N₁₀O₃ theory 682.2, found 683.3 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 10.45 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.58 (d, J = 1.8 Hz, 1H), 7.54 (dd, J = 8.6, 2.1 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 6.51 (dd, J = 8.9, 1.8 Hz, 1H), 6.44 (d, J = 1.4 Hz, 1H), 5.82 (d, J = 8.2 Hz, 1H), 4.58 (q, J = 9.4 Hz, 2H), 4.43 – 4.33 (m, 2H), 3.86 (t, J = 6.7 Hz, 2H), 3.82 (s, 3H), 3.70 (s, 2H), 3.68 – 3.60 (m, 1H), 3.16 – 3.07 (m, 2H), 2.72 (t, J = 6.7 Hz, 2H), 2.03 – 1.96 (m, 2H), 1.38 – 1.27 (m, 2H).

EXAMPLE S137.1 Synthesis of 1- (6- ((1- (5-chloro-4- ((1- (2-hydroxyethyl) -2-oxoindol-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (137)

Step 1 Synthesis of 1- (6- ((1- (5-chloro-4- ((1- (2-hydroxyethyl) -2-oxoindolin-5-yl) amino) pyrimidin-2-yl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedures 1 and 6 using 5-amino-1- (2-hydroxyethyl) indol-2-one and 1- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] hexahydropyrimidine-2, 4-dione as starting materials. LCMS C ₃₁H₃₃ClN₁₀O₄ theory 644.2, found 645.3 [M+H]⁺;¹H NMR (500 MHz, DMSO) δ 10.45 (s, 1H), 8.63 (s, 1H), 8.01 (s, 1H), 7.53 (s, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 6.99 (d, J = 8.5 Hz, 1H), 6.51 (dd, J = 8.9, 1.7 Hz, 1H), 6.44 (s, 1H), 5.82 (d, J = 8.3 Hz, 1H), 4.79 (bs, 1H), 4.44 – 4.32 (m, 2H), 3.86 (t, J = 6.7 Hz, 2H), 3.82 (s, 3H), 3.70 (t, J = 5.9 Hz, 2H), 3.68 – 3.60 (m, 1H), 3.57 (t, J = 5.8 Hz, 2H), 3.53 (s, 2H), 3.16 – 3.07 (m, 2H), 2.72 (t, J = 6.7 Hz, 2H), 2.04 – 1.96 (m, 2H), 1.38 – 1.26 (m, 2H).

EXAMPLE S138.Synthesis of 2- (4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) piperidin-1-yl) -4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidine-5-carbonitrile (138)

Step 1 Synthesis of 2- (4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) piperidin-1-yl) -4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidine-5-carbonitrile. The title compound was synthesized according to general procedures 1 and 6 using 2, 4-dichloropyrimidine-5-carbonitrile and 1- [ 1-methyl-6- (4-piperidylamino) indazol-3-yl ] hexahydropyrimidine-2, 4-dione, hydrochloride as starting materials. LCMS C ₃₁H₃₁N₁₁O₃ theory was 605.3, found to be 606.5 [M+H]⁺; ¹H NMR (500 MHz, DMSO) δ 10.45 (s, 1H), 9.28 (s, 1H), 8.45 (s, 1H), 8.40 (s, 1H), 7.52 – 7.40 (m, 2H), 7.28 (d, J = 8.8 Hz, 1H), 6.98 – 6.81 (m, 1H), 6.51 (dd, J = 8.9, 1.7 Hz, 1H), 6.45 (s, 1H), 5.84 (d, J = 8.2 Hz, 1H), 4.67 – 4.50 (m, 1H), 4.41 – 4.27 (m, 1H), 3.86 (t, J = 6.7 Hz, 2H), 3.82 (s, 3H), 3.72 – 3.62 (m, 1H), 3.54 (s, 2H), 3.26 – 3.15 (m, 2H), 3.10 (s, 3H), 2.72 (t, J = 6.7 Hz, 2H), 2.03 (d, J = 10.1 Hz, 2H), 1.32 (td, J= 13.6, 4.0 Hz, 2H).MS (ESI) [M+H]⁺.

EXAMPLE S139.1 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione (139)

Step 1 Synthesis of 1- (6-bromo-1-methyl-1H-indazol-3-yl) urea. To a stirred solution of 6-bromo-1-methyl-1H-indazol-3-amine (10 g,44.2 mmol) in acetonitrile (100 mL) was added trimethylsilane isocyanate (20.4 g,177 mmol) in a sealed tube. The reaction mixture was heated to 65 ℃ and stirred 16 h. The reaction mixture was cooled to 25 ℃ and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel with 10% methanol/DCM to give the title compound as a brown solid (4.0 g,13.7 mmol,30% yield).

Step 2 Synthesis of 1- (6-bromo-1-methyl-1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 1- (6-bromo-1-methyl-1H-indazol-3-yl) urea (4.0 g,13.67 mmol) in DMF (10 mL) at 0 ℃ under nitrogen was added methyl 3, 3-dimethoxypropionate (2.431 g,16.40 mmol). The reaction mixture was heated to 90 ℃ and stirred 3 h. The reaction mixture was cooled to room temperature and treated with ice-cold water to obtain a solid. The solid was filtered and washed with water to give the title compound as a yellow solid (4.1 g,4.63 mmol,33% yield). LCMS 320.9 [ M+H ] ⁺.

Step 3 Synthesis of 1- (6-bromo-1-methyl-1H-indazol-3-yl) -3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 1- (6-bromo-1-methyl-1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione (4.0 g,12.46 mmol) in DMF (20 mL) was added cesium carbonate (6.09 g,18.68 mmol) and 1- (bromomethyl) -4-methoxybenzene (3.76 g,18.68 mmol). The reaction mixture was slowly heated to 80 ℃ and stirred 3 h. The reaction mixture was poured into water and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude product. The crude product was purified by flash column chromatography on silica gel with 30% ethyl acetate/petroleum ether to give the title compound (2.05 g,3.58 mmol,28% yield).

Step 4 Synthesis of (3R, 4R) -4- ((3- (3- (4-methoxybenzyl) -2, 4-dioxo-3, 4-dihydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester. To a stirred solution of 1- (6-bromo-1-methyl-1H-indazol-3-yl) -3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione (500 mg,0.872 mmol) in 1, 4-dioxane (10 mL) was added cesium carbonate (710 mg,2.181 mmol) and tert-butyl (3 r,4 r) -4-amino-3-methylpiperidine-1-carboxylate (280 mg 1.30 mmol), which was degassed with nitrogen 10 min. CPhos-Pd-G3 (703 mg,0.872 mmol) was then added. The reaction mixture was heated to 90 ℃ and stirred 18 h. The reaction mixture was filtered through a celite pad and the celite pad was washed with ethyl acetate. The filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography on silica gel with 30% ethyl acetate/petroleum ether to give the title compound as a pale yellow solid (301 mg,0.426 mmol,48% yield).

Step 5 Synthesis of 1- (1-methyl-6- (((3R, 4R) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of (3 r,4 r) -4- ((3- (3- (4-methoxybenzyl) -2, 4-dioxo-3, 4-dihydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester (300 mg,0.42 mmol) in 1, 4-dioxane (10 mL) was added TFA (5.0 mL). The reaction mixture was heated to 80 ℃ and stirred 48 h. The reaction mixture was concentrated under reduced pressure and triturated with diethyl ether to give the title compound as a brown gummy solid (301 mg,0.426 mmol,48% yield). The crude product was used in the next step without further purification.

Step 6 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 1- (1-methyl-6- (((3 r,4 r) -3-methylpiperidin-4-yl) amino) -1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione (100 mg,0.172 mmol) in DMSO (2.0 mL) was added 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (50.3 mg,0.172 mmol) and DIPEA (0.15 mL,0.85 mmol) at 25 ℃. The reaction mixture was heated to 80 ℃ and stirred for 6 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC to give the title compound as an off-white solid (7.0 mg,0.01 mmol,5% yield). The preparation method comprises Rt 10.5 min, column X-select C18 (150X 19) mm,5.0 μm, 0.1% formic acid/acetonitrile in water, flow rate 15.0: 15.0 mL/min. LCMS C ₃₁H₃₁N₁₀O₃ theory 626.2, found 627.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 11.55 (s, 1H), 8.68 (s, 1H), 8.03 (s, 1H), 7.74 (d, J = 7.60 Hz, 1H), 7.57-7.52 (m, 2H), 7.23 (d, J = 8.80 Hz, 1H), 6.95 (d, J = 8.00 Hz, 1H), 6.58 (d, J = 8.0 Hz, 1H), 6.5 (s, 1H), 5.90 (d, J = 8.80 Hz, 1H), 5.71 (d, J = 7.60 Hz, 1H), 4.52-4.46 (m, 2H), 3.88 (s, 3H), 3.54 (s, 2H), 3.11 (s, 3H), 3.06-2.99 (m, 1H), 2.72-2.69 (m, 1H), 2.33-2.29 (m, 1H), 2.09-2.07 (m, 1H), 1.62-1.58 (m, 1H), 1.31-1.17 (m, 2H), 0.98 (d, J = 5.60 Hz, 3H).

EXAMPLE S140.1 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -5-fluoropyrimidine-2, 4 (1H, 3H) -dione (140)

Step 1 Synthesis of 5-fluoro-3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 5-fluoropyrimidine-2, 4 (1H, 3H) -dione (5.0 g,38.4 mmol) in DMF (100 mL) under nitrogen at 25℃was added triethylamine (5.36 mL,38.4 mmol). The reaction mixture was cooled to 0 ℃ and methyl chloroformate (2.98 mL,38.4 mmol) was added dropwise and stirred 3 h. Et ₃ N (8.04 ml,57.6 mmol) and 4-methoxybenzyl chloride (7.82 mL,57.7 mmol) were then added at 0 ℃. The reaction mixture was slowly warmed to 25 ℃ and stirred 6 h. The reaction mixture was treated with ice-cold water (100 mL) and extracted with ethyl acetate (2×100 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product. The crude product was dissolved in a 1:1 mixture of MeOH and DCM (50 mL) and 30% H ₂O₂ (4.32 mL,42.3 mmol) was added followed by 6.0N NaOH (0.1 mL) at 0 ℃. The reaction mixture was stirred 1h and treated with ice-cold water (100 mL) and extracted with dichloromethane (2 x100 mL). The combined organic layers were washed with water, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product. The crude product was purified by flash column chromatography on silica gel with 50% ethyl acetate/petroleum ether to give the title compound as an off-white solid (5.10 g,17.58 mmol,46% yield ）.¹H NMR (400 MHz, DMSO-d₆): δ 11.16 (d, J = 5.6 Hz, 1H), 7.87 (t, J = 6.0 Hz, 1H), 7.26-7.23 (m, 2H), 6.88-6.86 (m, 2H), 4.88 (s, 2H), 3.72 (s, 3H).

Step 2 Synthesis of 1- (6-bromo-1-methyl-1H-indazol-3-yl) -5-fluoro-3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 6-bromo-1-methyl-3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole (3.0 g,8.90 mmol) and molecular sieve (0.5 g) in DCM (40 mL) was added the compound 5-fluoro-3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione (5.06 g,17.80 mmol), et ₃ N (2.481 mL,17.80 mmol) and copper (II) acetate (1.617 g,8.90 mmol) at 25 ℃. The reaction mixture was stirred under oxygen 16 h. The reaction mixture was filtered through a celite pad and the filtrate was concentrated in vacuo to give the crude compound. The crude product was purified by flash column chromatography on silica gel with 35% -45% ethyl acetate/petroleum ether to give the title compound as an off-white solid (1.2 g,2.61 mmol,29% yield ）.¹H NMR (400 MHz, DMSO-d₆): δ 8.38 (d, J = 6.0 Hz, 1H), 8.11 (d, J = 1.2 Hz, 1H), 7.65 (dd, J = 0.4 Hz and 8.8 Hz, 1H), 7.37 (dd, j=1.2 Hz and 8.4 Hz, 1H), 7.33-7.30 (m, 2H), 6.91-6.88 (m, 2H), 5.00 (s, 2H), 4.07 (s, 3H), 3.73 (s, 3H).

Step 3 Synthesis of (3R, 4R) -4- ((3- (5-fluoro-3- (4-methoxybenzyl) -2, 4-dioxo-3, 4-dihydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methylpiperidine-1-carboxylic acid tert-butyl ester. A stirred solution of the compound 1- (6-bromo-1-methyl-1H-indazol-3-yl) -5-fluoro-3- (4-methoxybenzyl) pyrimidine-2, 4 (1H, 3H) -dione (1.10 g,2.395 mmol) and tert-butyl (3 r,4 r) -4-amino-3-methylpiperidine-1-carboxylate (513 mg,2.395 mmol) in 1, 4-dioxane (30.0 mL) was degassed with nitrogen 5 min. CPhos-Pd-G3 (97 mg,0.120 mmol) and Cs ₂CO₃ (1.171 g,3.59 mmol) were then added. The reaction mixture was heated to 100 ℃ and stirred 10 h. The reaction mixture was cooled to 25 ℃, treated with water (100 mL) and extracted with ethyl acetate (2×100 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product. The crude product was purified by flash column chromatography on silica gel with 80% -100% ethyl acetate/petroleum ether to give the title compound as an off-white solid (400 mg,0.621 mmol,26% yield). LCMS 593.2 (M+H), rt 3.467 min

Step 4 Synthesis of 5-fluoro-1- (1-methyl-6- (((3R, 4R) -3-methyl-1- (2, 2-trifluoroacetyl) -1l 4-piperidin-4-yl) amino) -1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione. CF ₃SO₃ H (0.375 mL,4.22 mmol) was added to a stirred solution of compound 6 (500 mg,0.844 mmol) in TFA (0.65 mL,8.44 mmol) at 25 ℃. The reaction mixture was heated to 50 ℃ and stirred 8 h. The reaction mixture was concentrated under vacuum to give the crude product. The crude compound was triturated with diethyl ether and dried under reduced pressure to give the title compound as a brown solid (400 mg,0.782 mmol,93% yield). The crude product was used in the next step without further purification. LCMS 372.7 [ M+H ] ⁺.

Step 5 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methylpiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -5-fluoropyrimidine-2, 4 (1H, 3H) -dione. To a stirred solution of 5-fluoro-1- (1-methyl-6- (((3 r,4 r) -3-methyl-1- (2, 2-trifluoroacetyl) -1l 4-piperidin-4-yl) amino) -1H-indazol-3-yl) pyrimidine-2, 4 (1H, 3H) -dione (100 mg,0.195 mmol- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (58.1 mg,0.195 mmol) in DMSO (2.0 mL) at 25 ℃ was added DIPEA (0.102 mL,0.586 mmol) in DMSO (2.0 mL). The reaction mixture was heated to 80 ℃ and stirred 4 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC to give the title compound as an off-white solid (30 mg,0.045 mmol,23% yield). LCMS C ₃₁H₃₀ClFN₁₀O₃ theory 644.2, found 645.2 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆): δ 12.05 (s, 1H), 8.69 (s, 1H), 8.20 (d, J = 6.4 Hz, 1H), 8.03 (s, 1H), 7.58 (s, 1H), 7.52(dd, J = 1.6 Hz and 8.8 Hz, 1H), 7.27 (d, j=8.8 Hz, 1H), 6.94 (d, j=8.4 Hz, 1H), 6.59 (dd, j=1.6 Hz and 9.2 Hz, 1H), 6.51 (s, 1H), 5.90 (d, J = 9.2 Hz, 1H), 4.49-4.46 (m, 2H), 3.88 (s, 3H), 3.54 (s, 2H), 3.14 (s, 3H), 3.11-3.00 (m, 1H), 2.73-2.67 (m, 1H), 2.09-2.07 (m, 1H), 1.56-1.63 (m, 1H), 1.24-1.15 (m, 1H), 0.98 (d, J = 6.4 Hz, 3H).

EXAMPLES S141-S144 Synthesis of Compounds 141-144

The examples in table 4 below were prepared according to the general procedure outlined in the table starting with their respective commercial starting materials and intermediates found in this document.

EXAMPLE S145.2 Synthesis of- ((3R, 4R) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile (145)

Step 1 Synthesis of tert-butyl (3R, 4R) -4- (((benzyloxy) carbonyl) amino) -3-hydroxypiperidine-1-carboxylate. To a stirred solution of tert-butyl (3R, 4R) -4-amino-3-hydroxypiperidine-1-carboxylate (2.5 g,11.6 mmol) in DCM (10 mL) at 0℃was added Cbz-Cl (1.97 g,11.6 mmol) and saturated aqueous sodium carbonate (10 mL). The reaction mixture was warmed to 25 ℃ and stirred 2 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude residue was purified by flash column chromatography on silica with 40% ethyl acetate/petroleum ether to give the title compound as an off-white solid (3.3 g,9.42 mmol,81% yield).

Step 2 synthesis of tert-butyl (3R, 4R) -4- (((benzyloxy) carbonyl) amino) -3-methoxypiperidine-1-carboxylate. To a stirred solution of silver (I) oxide (10.9 g,47.1 mmol) and tert-butyl (3R, 4R) -4- (((benzyloxy) carbonyl) amino) -3-hydroxypiperidine-1-carboxylate (3.3 g,9.42 mmol) in ACN (50 mL) was added methyl iodide (14.7 mL,235 mmol) dropwise. The reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (3×50 mL). The filtrate was concentrated under reduced pressure to give a crude product. The crude residue was purified by flash column chromatography on silica with 25% ethyl acetate/petroleum ether to give the title compound as a colorless liquid (3.1 g,8.45 mmol,90% yield).

Step 3 synthesis of (3R, 4R) -4-amino-3-methoxypiperidine-1-carboxylic acid tert-butyl ester. To a stirred solution of tert-butyl (3R, 4R) -4- (((benzyloxy) carbonyl) amino) -3-methoxypiperidine-1-carboxylate (3.0 g,8.23 mmol) in methanol (20 mL) was added 10% palladium on carbon (1.0 g) under nitrogen. The reaction mixture was stirred at 25 ℃ at a hydrogenation pressure of 1 atm at 3 h. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (200 mL). The filtrate was concentrated under reduced pressure to give the title compound as a crude product (1.7 g,7.38 mmol,90% yield). The crude product was used in the next step without further purification.

Step 4 Synthesis of tert-butyl (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylate. A stirred solution of (3R, 4R) -4-amino-3-methoxypiperidine-1-carboxylic acid tert-butyl ester (200 mg,0.87 mmol) and 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (435 mg,0.87 mmol) in toluene (10 mL mmol) was degassed 10 min. NaO ^tBu（167 mg,1.74 mmol）、Pd₂dba₃ (80 mg,0.09 mmol) and CPhos (114 mg,2.26 mmol) were then added at 25 ℃. The reaction mixture was heated to 100 ℃ and stirred 12 h. The reaction mixture was poured into water (150 mL) and diluted with ethyl acetate (150 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3×150 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude residue was purified by flash column chromatography on silica with 25% ethyl acetate/petroleum ether to give the title compound as a light brown solid (485 mg,0.66 mmol,76% yield).

Step 5 Synthesis of tert-butyl (3R, 4R) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylate. To a stirred solution of (3 r,4 r) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester (485 mg,0.66 mmol) and trifluoroacetic acid (0.05 mL,0.66 mmol) in ethanol (4 mL) and THF (4 mL) was added 20% palladium on carbon hydroxide (150 mg) under nitrogen. The reaction mixture was stirred at 25 ℃ at a hydrogenation pressure of 1 atm at 3 h. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (200 mL). The filtrate was concentrated under reduced pressure to give the title compound as a crude product (300 mg,0.42 mmol,64% yield). The crude product was used in the next step without further purification.

Step 6 Synthesis of 3- (6- (((3R, 4R) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedure 5 using (3 r,4 r) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester (250 mg,0.53 mmol) as starting material.

Step 7 Synthesis of 2- ((3R, 4R) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile. The synthesis of the title compound was completed using general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (200 mg,0.38 mmol) and 4- ((1-methyl-2-oxoindol-5-yl) amino) -2- (methylsulfonyl) pyrimidine-5-carbonitrile (149 mg,0.38 mmol). The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (7.7 mg,0.012 mmol,8.23% yield ）.MS (ESI) m/z 635.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 9.29 (s, 1H), 8.39 (d, J = 11.20 Hz, 1H), 7.50 (d, J = 11.20 Hz, 2H), 7.44 (d, J = 7.60 Hz, 1H), 6.93 (d, J = 8.40 Hz, 1H), 6.58 (d, J = 8.80 Hz, 1H), 6.47 (s, 1H), 5.90 (s, 1H), 4.21- 4.07 (m, 2H), 4.21- 4.07 (m, 3H), 3.92 - 3.83 (m, 1H), 3.82 (s, 3H), 3.81 - 3.60 (m, 2H), 3.54 (s, 3H), 3.53 - 3.48 (m, 1H), 3.10 (m, 3H), 2.62 (d, J = 8.40 Hz, 2H), 2.30 - 2.00 (m, 3H), 1.36 (brs, 1H).

Example S146.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (146)

Step 1 Synthesis of (3R, 4R) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester. A solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (1750 mg,3.50 mmol), tert-butyl (3R, 4R) -4-amino-3-methoxypiperidine-1-carboxylate (1007 mg,4.37 mmol) and sodium tert-butoxide (504 mg,5.25 mmol) in dioxane (35 mL) was degassed with nitrogen 15min before Josiphos Pd G3 (323 mg,0.350 mmol) was added. The reaction mixture was then heated at 100 ℃ to 16 h. The mixture was then cooled to room temperature and diluted with saturated sodium bicarbonate and extracted with EtOAc (3×). The combined organics were dried over magnesium sulfate and filtered through celite, then concentrated and purified by silica gel chromatography using 0to 100% EtOAc in hexanes to provide the title compound （1.25 g,1.92 mmol）.¹H NMR (500 MHz, CDCl₃) δ ppm 7.92 (d, J = 7.99 Hz, 1H), 7.54 - 7.29 (m, 11H), 7.28 - 7.25 (m, 1H), 6.51 (d, J = 8.11 Hz, 1H), 6.47 - 6.41 (m, 2H), 5.49 (s, 2H), 5.41 (s, 2H), 4.00 (s, 4H), 3.96 - 3.73 (m, 2H), 3.49 (s, 4H), 3.22 (td, J = 7.60, 3.87 Hz, 2H), 2.30 - 2.19 (m, 1H), 1.51 (s, 9H).

Step 2 Synthesis of 3- (6- (((3R, 4R) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride. The synthesis of the title compound was completed using general procedures 4 and 5 using (3 r,4 r) -4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester as starting material.

Step 3 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The synthesis of the title compound was completed using general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione hydrochloride (65 mg,0.159 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (46 mg,0.159 mmol). The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound as a white solid (7.7 mg,0.012 mmol,8.23% yield ）.MS (ESI) m/z 644.6 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 8.67 (s, 1H), 8.03 (s, 1H), 7.64 - 7.47 (m, 2H), 7.33 (d, J = 8.8 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.56 (dd, J = 1.7, 8.8 Hz, 1H), 6.46 (d, J = 1.2 Hz, 1H), 5.85 (d, J = 8.0 Hz, 1H), 4.29 (br d, J = 11.4 Hz, 1H), 4.18 (dd, J = 5.1, 8.8 Hz, 1H), 4.11 - 3.99 (m, 1H), 3.82 (s, 3H), 3.67 - 3.56 (m, 1H), 3.54 (s, 2H), 3.42 - 3.35 (m, 1H), 3.30 (s, 3H), 3.17 (dt, J = 3.9, 7.4 Hz, 1H), 3.10 (s, 3H), 2.66 - 2.57 (m, 2H), 2.32 - 2.21 (m, 1H), 2.19 - 2.11 (m, 1H), 2.07 - 1.99 (m, 1H), 1.42 - 1.31 (m, 1H).

EXAMPLE S147 Synthesis of relative-3- (6- (((3R, 4S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (trifluoromethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione, isomer 1 (147).

Step 1 Synthesis of 4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylic acid tert-butyl ester. 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (300 mg,0.600 mmol), cesium carbonate (391 mg,1.199 mmol), 4-amino-3- (trifluoromethyl) piperidine-1-carboxylic acid tert-butyl ester (241 mg,0.899 mmol) and XPhos Pd G3 (50.7 mg,0.060 mmol) were added to a 1-dram vial equipped with a stirring bar and purged with nitrogen gas 1 min. Next, dioxane (5995 μl) was added and the reaction mixture was stirred at 70 ℃ overnight. LCMS indicated significant product formation. The reaction was cooled to room temperature and poured into a separatory funnel with water, saturated potassium carbonate and dichloromethane. The aqueous layer was extracted 3X with dichloromethane. The combined organic layers were dried over magnesium sulfate, filtered and concentrated. The mixture was purified with 0-50% EA/hexane and 1% MeOH. Fractions containing the clean product were combined and lyophilized to give the title compound tert-butyl 4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylate (354 mg,0.515 mmol,86% yield) as a yellow solid. Enantiomers were separated by chiral SFC use (isocratic: 25B, where B is 0.1% FA in methanol; column: CHIRALPACK IC, flow rate: 2 ml/min). This gives the relative enantiomer tert-butyl ester of relative- (3R, 4S) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylate, isomer 1 (90.1 mg,24.5% yield) and the relative- (3R, 4S) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylate, isomer 2 (87.4 mg,24.5% yield). LC MS shows 688.311 at t=3.89 (m+1= 688.76). ¹ H NMR (chloroform -d, 400 MHz) δ 7.8-8.0 (m, 1H), 7.3-7.5 (m, 8H), 6.4-6.6 (m, 1H), 6.3-6.4 (m, 2H), 5.3-5.5 (m, 4H), 4.0-4.1 (m, 3H), 4.0-4.0 (m, 3H), 3.5-3.7 (m, 2H), 2.17 (s, 3H), 2.0-2.1 (m, 1H), 1.6-1.8 (m, 2H), 1.5-1.5 (m, 9H).)

Step 2 Synthesis of isomer 1 from tert-butyl relative- (3R, 4S) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylate. 4- ((3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylic acid tert-butyl ester, isomer 1 (90.1 mg,0.131 mmol) and MeOH (1310 μl) were added to a 40 mL vial equipped with a stirring bar. The reaction was purged with nitrogen and then palladium on carbon (27.9 mg,0.013 mmol) was added. The vial was purged again with nitrogen and then a hydrogen balloon was added. The vials were heated to 50 ℃ for 15 hours. The mixture was filtered through celite. The filtrate was concentrated to give the title compound (56 mg,84% yield). LCMS showed 510.2 at t=2.86 and 2.93 (m+1=510.2). ¹ H NMR (chloroform -d, 500 MHz) δ 8.0-8.2 (m, 1H), 7.4-7.6 (m, 1H), 6.5-6.6 (m, 1H), 6.3-6.4 (m, 1H), 4.2-4.3 (m, 1H), 4.0-4.2 (m, 2H), 3.9-4.0 (m, 4H), 3.8-3.8 (m, 1H), 3.4-3.5 (m, 1H), 2.9-3.1 (m, 1H), 2.6-2.8 (m, 2H), 2.4-2.6 (m, 1H), 2.3-2.4 (m, 1H), 2.0-2.1 (m, 1H), 1.9-1.9 (m, 1H), 1.7-1.9 (m, 1H), 1.50 (s, 10H).)

Step 3 Synthesis of 3- (1-methyl-6- (((3R, 4S) -3- (trifluoromethyl) piperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione, isomer 1. A round bottom flask was charged with the relative tert-butyl- (3R, 4S) -4- ((3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) amino) -3- (trifluoromethyl) piperidine-1-carboxylate (56.0 mg,0.110 mmol) in DCM (1099. Mu.L). HCl (55.0 μl,0.220 mmol) was then added and the solution was stirred at room temperature for 1 hour. Next, the solution was concentrated to give the title compound as HCl salt as a white solid (49 mg,0.110 mmol,100% yield), which was used in the next step without further purification. LCMS showed 410.2 at t=141 and 1.53 (m+1=410.2).

Step 4 Synthesis of relative-3- (6- (((3R, 4S) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (trifluoromethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione, isomer 1. To a stirred solution of 3- (1-methyl-6- (((3 r,4 s) -3- (trifluoromethyl) piperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione, isomer 1 (28 mg,0.075 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (28.6 mg,0.075 mmol) in DMSO (1 mL) was added DIPEA (0.053 mL,0.302 mmol). The reaction was stirred at 80 ℃ for 4 hours. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound (3.2 mg,4.69 μmol,4.26% yield). ¹ H NMR (chloroform -d, 400 MHz) δ 8.0-8.1 (m, 1H), 7.8-7.9 (m, 1H), 7.4-7.6 (m, 3H), 6.9-7.0 (m, 1H), 6.7-6.8 (m, 1H), 6.5-6.7 (m, 1H), 6.2-6.4 (m, 1H), 4.5-4.6 (m, 1H), 4.1-4.3 (m, 4H), 3.91 (s, 3H), 3.6-3.7 (m, 1H), 3.52 (s, 3H), 3.22 (s, 3H), 2.9-3.1 (m, 1H), 2.6-2.8 (m, 2H), 2.3-2.6 (m, 2H), 2.0-2.2 (m, 1H), 2.0-2.0 (m, 1H), 1.7-1.8 (m, 1H).LC MS at t=2.49 (m+1= 682.1) showed 682.3.

EXAMPLE S148.Synthesis of 3- (6- (((3S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (trifluoromethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (148)

Step 1 Synthesis of 3- (6- (((3S, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3- (trifluoromethyl) piperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione, isomer 2. The title compound was synthesized in analogy to example S147 using the second eluting isomer from step 1 of example S147. To a stirred solution of 3- (1-methyl-6- (((3 s,4 r) -3- (trifluoromethyl) piperidin-4-yl) amino) -1H-indazol-3-yl) piperidine-2, 6-dione, isomer 2 (56 mg,0.137 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (40.0 mg,0.137 mmol) in DMSO (1 mL) was added N-ethyl-N-isopropyl-2-amine (0.096 mL,0.547 mmol). The reaction was stirred at 80 ℃ for 4 hours. The reaction mixture was filtered and purified by reverse phase semi-preparative HPLC (10% -100% acetonitrile + 0.1% formic acid in water (0.1% formic acid), via 30 min). Fractions containing clean product were combined and lyophilized to give the title compound (21.7 mg,0.031 mmol,23.03% yield ）.¹H NMR (CDCl₃, 400 MHz) δ 8.0-8.1 (m, 1H), 7.8-7.9 (m, 1H), 7.4-7.6 (m, 3H), 6.9-7.0 (m, 1H), 6.7-6.8 (m, 1H), 6.5-6.7 (m, 1H), 6.2-6.4 (m, 1H), 4.5-4.6 (m, 1H), 4.1-4.3 (m, 4H), 3.91 (s, 3H), 3.6-3.7 (m, 1H), 3.52 (s, 3H), 3.22 (s, 3H), 2.9-3.1 (m, 1H), 2.6-2.8 (m, 2H), 2.3-2.6 (m, 2H), 2.0-2.2 (m, 1H), 2.0-2.0 (m, 1H), 1.7-1.8 (m, 1H).LCMS [M+1 = 682.1] showed 682.3.

Example S149.Synthesis of 3- (6- ((4R, 5S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -5-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (149)

Step 1 (S) - (3- (2, 2-dimethyl-4, 6-dioxo-1, 3-dioxan-5-yl) -2-methyl-3-oxopropyl) carbamic acid tert-butyl ester synthesis. To a stirred solution of (S) -3- ((tert-butoxycarbonyl) amino) -2-methylpropanoic acid (10.00 g,46.7 mmol) and 2, 2-dimethyl-1, 3-dioxane-4, 6-dione (7.41 g,51.4 mmol) in DCM (160 mL) was added EDC (13.44 g,70.1 mmol) and DMAP (8.57 g,70.1 mmol). The reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was diluted with DCM (300 mL), washed with 10% aqueous sodium bisulfate (3×100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude title compound as a yellow solid (16.2 g,27.5 mmol,59% yield). The crude product was used in the next step without further purification.

Step 2 synthesis of (S) -5-methyl-2, 4-dioxopiperidine-1-carboxylic acid tert-butyl ester. A solution of (S) - (3- (2, 2-dimethyl-4, 6-dioxo-1, 3-dioxan-5-yl) -2-methyl-3-oxopropyl) carbamic acid tert-butyl ester (16.2 g,27.5 mmol) in ethyl acetate (150 mL) was refluxed at 80℃and stirred for 8 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give the crude compound. The crude material was purified by flash column chromatography on silica with 50% ethyl acetate/petroleum ether to give the title compound (6.92 g,27.4 mmol, quantitative yield) as a white solid.

Step 3 synthesis of (S) -5-methylpiperidine-2, 4-dione. Trifluoroacetic acid (21.1 ml,274 mmol) was added dropwise to a stirred solution of tert-butyl (S) -5-methyl-2, 4-dioxopiperidine-1-carboxylate (6.92 g,27.4 mmol) in DCM (100 mL) at 0 ℃. The resulting solution was warmed to 25 ℃ and stirred 16 h. The reaction mixture was concentrated under reduced pressure, dissolved in acetonitrile (100 mL), and basified by solid sodium bicarbonate until the pH of the solution was greater than 8. The mixture was stirred for an additional hour, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude title compound as a yellow oil (8.5 g,26.9 mmol,98% yield). The crude product was used in the next step without further purification.

Step 4 synthesis of (S) -4- (benzyl (methyl) amino) -5-methyl-5, 6-dihydropyridin-2 (1H) -one. To a stirred solution of (S) -5-methylpiperidine-2, 4-dione (8.5 g,26.9 mmol) and N-methyl-1-phenylmethylamine (6.52 g,53.8 mmol) in MeOH (150 mL) was added AcOH (2.3 mL,40.4 mmol). The reaction mixture was stirred at 60 ℃ for 16 h. The reaction mixture was concentrated under reduced pressure to give the title compound, which was used without further purification (20 g,26.4 mmol,98% yield).

Step 5 synthesis of (4R, 5S) -5-methyl-4- (methylamino) piperidin-2-one. To a solution of (S) -4- (benzyl (methyl) amino) -5-methyl-5, 6-dihydropyridin-2 (1H) -one (20 g,26.4 mmol) in methanol (150 mL) was added platinum (IV) oxide (3.0 g). The reaction mixture was stirred at 25 ℃ at 7 atm hydrogenation pressure 72 h. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (3 x 100 mL). The filtrate was concentrated under reduced pressure. This material was purified by preparative HPLC to give the title compound as a white solid (3.8 g,26.3 mmol,99% yield). The preparative HPLC method was Rt 5.4 min, column XSelect C18 (300X 30) mm,10.0 μm, 0.1% TFA in water/acetonitrile, flow rate 20.0 mL/min.

Step 6 synthesis of tert-butyl methyl ((4R, 5S) -5-methyl-2-oxopiperidin-4-yl) carbamate. To a stirred solution of (4R, 5S) -5-methyl-4- (methylamino) piperidin-2-one (3.8 g,26.3 mmol) in THF (38 mL) were added Et ₃ N (9.2 mL,65.8 mmol) and boc-anhydride (6.32 g,28.9 mmol) in sequence. The reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was diluted with ethyl acetate (30 mL) and quenched with water (20 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound (1.32 g,4.15 mmol,16% yield) which was used without further purification.

Step 7 synthesis of (((4R, 5S) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -5-methyl-2-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester. A stirred solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (1.32 g,2.64 mmol) and methyl ((4R, 5S) -5-methyl-2-oxopiperidin-4-yl) carbamic acid tert-butyl ester (0.7 g,2.20 mmol) in toluene (25 mL) was degassed with nitrogen gas 5 min. Cesium carbonate (2.87 g,8.80 mmol), pd ₂(dba)₃ (0.20 g,0.22 mmol) and xantphos (0.51 g,0.88 mmol) were added. The reaction mixture was heated to 120 ℃ and stirred 16 h. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (50 mL) and quenched with water (50 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with water (3×50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound which was purified by preparative HPLC to give the title compound as a pale yellow solid (0.86 g,1.23 mmol,56% yield). The preparative HPLC method was Rt 7.4 min, column XSelect C18 (300X 30) mm,10.0 μm, 0.1% TFA in water/acetonitrile, flow rate 20.0 mL/min.

Step 8 synthesis of (((4R, 5S) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -5-methyl-2-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester. To a stirred solution of ((4 r,5 s) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -5-methyl-2-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (860 mg,1.23 mmol) and trifluoroacetic acid (0.10 mL,1.23 mmol) in ethanol (7.5 mL) and THF (7.5 mL) was added 20% palladium on carbon hydroxide (400 mg) under nitrogen. The reaction mixture was stirred at 50 ℃ under 4 atm hydrogenation pressure 4 h. The reaction mixture was filtered through a pad of celite and the pad was washed with ethanol/THF (1:1, 200 mL). The filtrate was concentrated under reduced pressure to give the title compound as a crude product (510 mg,0.70 mmol,57% yield). The crude product was used in the next step without further purification.

Step 9 Synthesis of 3- (6- ((4R, 5S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -5-methyl-2-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 5 and 6 using ((4 r,5 s) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -5-methyl-2-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (400 mg,0.55 mmol) as starting material. LCMS [ M+H ] ⁺ found 656.2;¹H NMR (400 MHz, DMSO-d₆): δ 10.92 (s, 1H), 8.67 (s, 1H), 8.05 (s, 1H), 7.70 (d, J = 8.40 Hz, 1H), 7.58-7.52 (m, 3H), 6.99-6.95 (m, 2H), 5.05 (brs, 1H), 4.41-4.37 (m, 1H), 3.99 (s, 3H), 3.70-3.65 (m, 1H), 3.57 (s, 3H), 3.33-3.25 (m, 3H), 3.08 (s, 3H), 2.83-2.63 (m, 5H), 2.40-2.34 (m, 1H), 2.21-2.17 (m, 1H), 0.99 (d, J = 6.80 Hz, 3H).

EXAMPLE S150.Synthesis of 3- (6- ((2R, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (150)

Step 1 Synthesis of tert-butyl methyl ((4R, 5S) -5-methyl-2-oxopiperidin-4-yl) carbamate. The title compound was synthesized according to the procedure of example S149, steps 1-6 using (R) -3- ((tert-butoxycarbonyl) amino) butyric acid as the original starting material.

Step 2 Synthesis of (((2R, 4S) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester. To a stirred solution of 3- (2, 6-bis (benzyloxy) pyridin-3-yl) -6-bromo-1-methyl-1H-indazole (4.13 g,8.25 mmol) and tert-butyl methyl ((4R, 5S) -5-methyl-2-oxopiperidin-4-yl) carbamate (2.0 g,8.25 mmol) in dioxane (40 mL) was added potassium phosphate (5.26 g,49.5 mmol), cuI (4.72 g,24.8 mmol) and DMEDA (4.37 g,49.5 mmol). The reaction mixture was heated to 120 ℃ and stirred 48 h. The reaction mixture was cooled to room temperature, quenched with saturated aqueous ammonium chloride (100 mL) and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with water (3×100 mL) and brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound which was purified by preparative HPLC to give the title compound as an off-white solid (0.41 g,0.62 mmol,8% yield, peak 1). The preparative HPLC method was Rt 15.0 min and 15.4 min, column YMC 18 Phenyl (250 x 21) mm,5.0 μm, 0.1% TFA in water/acetonitrile, flow rate 15.0 mL/min.

Step 3 Synthesis of (((2R, 4S) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester. To a stirred solution of ((2 r,4 s) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (410 mg,0.62 mmol, peak 1) and trifluoroacetic acid (0.05 mL,0.62 mmol) in ethanol (3 mL) and THF (3 mL) under nitrogen was added 20% palladium on carbon hydroxide (250 mg). The reaction mixture was stirred at 50 ℃ under 4 atm hydrogenation pressure 4 h. The reaction mixture was filtered through a pad of celite and the pad was washed with ethanol/THF (1:1, 150 mL). The filtrate was concentrated under reduced pressure to give the title compound as a crude product (300 mg,0.61 mmol,99% yield). The crude product was used in the next step without further purification.

Step 4 Synthesis of 3- (6- ((2R, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 5 and 6 using ((2 r,4 s) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (300 mg,0.61 mmol) as starting material. LCMS [ M+H ] ⁺ found 656.2;¹H NMR (400 MHz, DMSO-d₆): δ 10.92 (s, 1H), 8.68 (s, 1H), 8.05 (s, 1H), 7.70 (d, J = 8.80 Hz, 1H), 7.60 (s, 1H), 7.51 - 7.46 (m, 2H), 6.93 (d, J = 8.40 Hz, 1H), 6.85 (s, 1H), 5.00 (brs, 1H), 4.42 - 4.38 (m, 1H), 4.05 - 4.00 (m, 1H), 3.99 (s, 3H), 3.54 - 3.52 (m, 2H), 3.09 (s, 3H), 2.99 (s, 3H), 2.66 - 2.64 (m, 3H), 2.45 - 2.41 (m, 2H), 2.22 - 2.08 (m, 2H), 1.98 - 1.95 (m, 1H), 0.93 (d, J = 6.00 Hz, 3H).

EXAMPLE S151.3 Synthesis of 3- (6- ((2R, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (151)

Step 1 Synthesis of (((2R, 4R) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester. To a stirred solution of ((2 r,4 r) -1- (3- (2, 6-bis (benzyloxy) pyridin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (170 mg,0.26 mmol, peak 2 from example S150 step 2) and trifluoroacetic acid (0.02 mL,0.26 mmol) in ethanol (1 mL) and THF (1 mL) under nitrogen was added 20% palladium on carbon hydroxide (100 mg). The reaction mixture was stirred at 50 ℃ under 4 atm hydrogenation pressure 4 h. The reaction mixture was filtered through a pad of celite and the pad was washed with ethanol/THF (1:1, 50 mL). The filtrate was concentrated under reduced pressure to give the title compound as a crude product (135 mg,0.26 mmol,99% yield). The crude product was used in the next step without further purification.

Step 2 Synthesis of 3- (6- ((2R, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized according to general procedures 5 and 6 using ((2 r,4 r) -1- (3- (2, 6-dioxopiperidin-3-yl) -1-methyl-1H-indazol-6-yl) -2-methyl-6-oxopiperidin-4-yl) (methyl) carbamic acid tert-butyl ester (135 mg,0.26 mmol) as starting material. LCMS [ M+H ] ⁺ found 656.2;¹H NMR (400 MHz, DMSO-d₆): δ 10.91 (s, 1H), 8.74 (s, 1H), 8.05 (s, 1H), 7.71 (d, J = 8.80 Hz, 1H), 7.56 (s, 1H), 7.46 (s, 2H), 6.97 (d, J = 8.40 Hz, 1H), 6.89 (d, J = 7.60 Hz, 1H), 5.14 (brs, 1H), 4.42 - 4.38 (m, 1H), 4.13 - 4.10 (m, 1H), 4.01 (s, 3H), 3.57 (s, 2H), 3.14 (s, 3H), 2.98 (s, 3H), 2.61 - 2.60 (m, 4H), 2.44 - 2.33 (m, 2H), 2.23 - 2.17 (m, 1H), 1.81 - 1.79 (m, 1H), 0.98 (s, 3H).

EXAMPLE S152.3 Synthesis of 3- (6- ((2S, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (152)

Step 1 Synthesis of 3- (6- ((2S, 4R) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized in analogy to example S150 using (R) -3- ((tert-butoxycarbonyl) amino) butanoic acid (10.00 g,49.2 mmol) as starting material. LCMS [ M+H ] ⁺ found 656.2;¹H NMR (400 MHz, DMSO-d₆): δ 10.91 (s, 1H), 8.67 (s, 1H), 8.05 (s, 1H), 7.71 - 7.69 (m, 1H), 7.61 (s, 1H), 7.51 - 7.47 (m, 2H), 6.95 - 6.86 (m, 2H), 5.11 - 5.10 (m, 1H), 4.41 - 4.38 (m, 1H), 3.99 (s, 3H), 3.55 - 3.52 (m, 2H), 3.13 (s, 3H), 3.03 (s, 3H), 2.80 - 2.60 (m, 3H), 2.49 - 2.38 (m, 3H), 2.32 - 2.10 (m, 2H), 2.02 – 1.92 (m, 1H), 0.93 (d, J = 6.4 Hz, 3H).

EXAMPLE S153.Synthesis of 3- (6- ((2S, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione (153)

Step 1 Synthesis of 3- (6- ((2S, 4S) -4- ((5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) (methyl) amino) -2-methyl-6-oxopiperidin-1-yl) -1-methyl-1H-indazol-3-yl) piperidine-2, 6-dione. The title compound was synthesized in analogy to example S151. LCMS [ M+H ] ⁺ found 656.2;¹H NMR (400 MHz, DMSO-d₆): δ 10.91 (s, 1H), 8.72 - 8.68 (m, 1H), 8.05 (d, J = 2.0 Hz, 1H), 7.71 (dd, J = 4.40, 8.60 Hz, 1H), 7.60 (s, 1H), 7.56 - 7.46 (m, 2H), 6.98 - 6.88 (m, 2H), 5.21 -5.10 (m, 1H), 4.39 - 4.35 (m, 1H), 4.19 - 4.11 (m, 1H), 4.00 (d, J = 4.80 Hz, 3H), 3.55 - 3.52 (m, 2H), 3.13 (d, J = 17.60 Hz, 3H), 3.03 (d, J = 4.00 Hz, 3H), 2.80 - 2.67 (m, 2H), 2.62 - 2.60 (m, 1H), 2.47 - 2.38 (m, 2H), 2.30 - 2.28 (m, 2H), 2.05 - 2.00 (m, 1H), 1.02 (d, J = 6.40 Hz, 3H).

EXAMPLE S154.Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (154)

Step 1 Synthesis of (3R, 4R) -4- ((3- (5- (tert-butoxy) -2-cyano-5-oxopenta-2-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester. A stirred solution of tert-butyl 4- (6-bromo-1-methyl-1H-indazol-3-yl) -4-cyanovalerate (300 mg,0.77 mmol) and tert-butyl (3R, 4R) -4-amino-3-methoxypiperidine-1-carboxylate (176 mg,0.77 mmol) in toluene (10 mL) was degassed 5 min. NaO ^tBu（110 mg,1.15 mmol）、Pd₂dba₃ (70.0 mg,0.08 mmol) and BINAP (47.6 mg,0.08 mmol) were then added at 25 ℃. The reaction mixture was heated to 100 ℃ and stirred 6 h. The reaction mixture was quenched with water (20 mL) and diluted with ethyl acetate (50 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude residue was purified by flash column chromatography on silica with 100% ethyl acetate/petroleum ether to give the title compound as an off-white solid (300 mg,0.41 mmol,54% yield).

Step 2 Synthesis of tert-butyl (3R, 4R) -3-methoxy-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidine-1-carboxylate. To a stirred solution of (3R, 4R) -4- ((3- (5- (tert-butoxy) -2-cyano-5-oxopent-2-yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidine-1-carboxylic acid tert-butyl ester (300 mg,0.55 mmol) in acetic acid (5 mL) was added sulfuric acid (0.03 mL,0.55 mmol). The reaction mixture was heated to 60 ℃ and stirred 6 h. The reaction mixture was concentrated under reduced pressure and the resulting crude mixture was dissolved in acetonitrile (5 mL) followed by addition of DIPEA (0.27 mL,1.56 mmol) and BOC-anhydride (0.15 mL,0.62 mmol). The reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was quenched with water (20 mL) and diluted with ethyl acetate (30 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude residue was purified by flash column chromatography on silica with 100% ethyl acetate/petroleum ether to give the title compound as an off-white solid (180 mg,0.27 mmol, 49% yield over 2 steps).

Step 3 Synthesis of 3- (6- (((3R, 4R) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione. The title compound was synthesized according to general procedure 5 using (3 r,4 r) -3-methoxy-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidine-1-carboxylic acid tert-butyl ester (180 mg,0.37 mmol) as starting material.

Step 4 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione. The title compound was synthesized according to general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (100 mg,0.20 mmol) and 5- ((5-chloro-2-fluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (59 mg,0.20 mmol) as starting material. LCMS [ M+H ] ⁺ found 658.3;¹H NMR (400 MHz, DMSO-d6): δ 10.74 (s, 1H), 9.00 (s, 1H), 8.08 (s, 1H), 7.53 - 7.44 (m, 3H), 6.96 - 6.94 (m, 1H), 6.58 - 6.56 (m, 1H), 6.49 - 6.47 (m, 1H), 6.13 - 5.61 (m, 1H), 4.20 (br d, J = 12.5 Hz, 2H), 4.02 - 3.91 (m, 2H), 3.81 (s, 3H), 3.65 - 3.59 (m, 1H), 3.47 - 3.34 (m, 2H), 3.29 (s, 3H), 3.23 - 3.18 (m, 1H), 3.11 (s, 3H), 2.48 - 2.45 (m, 1H), 2.42 - 2.39 (m, 1H), 2.13 - 1.99 (m, 2H), 1.59 (s, 3H), 1.44 - 1.31 (m, 1H).

EXAMPLE S155.2 Synthesis of- ((3R, 4R) -3-methoxy-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile (155)

Step 1 Synthesis of 2- ((3R, 4R) -3-methoxy-4- ((1-methyl-3- (3-methyl-2, 6-dioxopiperidin-3-yl) -1H-indazol-6-yl) amino) piperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile. The title compound was synthesized according to general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (150 mg,0.30 mmol) and 2-fluoro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile (103 mg,0.30 mmol) as starting materials. LCMS [ M+H ] ⁺ found 649.2;¹H NMR (400 MHz, DMSO-d6): δ 10.77 (s, 1H), 9.30 (s, 1H), 8.40 (br s, 1H), 7.48 - 7.42 (m, 3H), 6.96 - 6.91 (m, 1H), 6.58 (br d, J = 8.0 Hz, 1H), 6.46 (s, 1H), 4.14 - 4.04 (m, 1H), 3.82 - 3.77 (m, 3H), 3.67 - 3.59 (m, 2H), 3.58 - 3.49 (m, 5H), 3.23 (br s, 2H), 3.10 (s, 3H), 2.58 - 2.53 (m, 2H), 2.48 - 2.45 (m, 1H), 2.43 - 2.37 (m, 1H), 2.13 - 1.98 (m, 2H), 1.59 (s, 3H), 1.45 - 1.37 (m, 1H).

Example S156.1 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (156)

Step 1 Synthesis of tert-butyl (3R, 4R) -3-methoxy-4- ((3- (3- (4-methoxybenzyl) -2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) piperidine-1-carboxylate. A stirred solution of 1- (6-bromo-1-methyl-1H-indazol-3-yl) -3- (4-methoxybenzyl) dihydropyrimidine-2, 4 (1H, 3H) -dione (1.00 g,2.26 mmol) and tert-butyl (3R, 4R) -4-amino-3-methoxypiperidine-1-carboxylate (0.52 g,2.26 mmol) in toluene (3 mL) was degassed for 10 min. Cs ₂CO₃ (1.47G, 4.51 mmol) and Pd-Ruphos-G3 (0.19G, 0.23 mmol) were then added at 25 ℃. The reaction mixture was heated to 120 ℃ and stirred 12 h. The reaction mixture was poured into water (150 mL) and diluted with ethyl acetate (150 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3×150 mL). The combined organic layers were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude residue was purified by flash column chromatography on silica with 70% ethyl acetate/petroleum ether to give the title compound as a pale yellow solid (0.80 g,1.27 mmol,56% yield).

Step 2 Synthesis of 1- (6- (((3R, 4R) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedure 5 using (3 r,4 r) -3-methoxy-4- ((3- (3- (4-methoxybenzyl) -2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) piperidine-1-carboxylic acid tert-butyl ester (800 mg,1.27 mmol) as starting material.

Step 3 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (150 mg,0.30 mmol) and 2-fluoro-4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile (103 mg,0.30 mmol) as starting materials. LCMS [ M+H ] ⁺ found 634.2;¹H NMR (400 MHz, DMSO-d6): δ 10.45 (s, 1H), 8.68 (s, 1H), 8.02 (s, 1H), 7.54 - 7.50 (m, 2H), 7.28 (d, J = 8.80 Hz, 1H), 6.93 (d, J = 8.40 Hz, 1H), 6.55 (dd, J = 1.60, 8.80 Hz, 1H), 6.45 (d, J = 1.20 Hz, 1H), 5.90 (d, J = 8.00 Hz, 1H), 4.32 (d, J = 10.80 Hz, 1H), 4.07 (brs, 1H), 3.86 (t, J = 6.80 Hz, 2H), 3.81 (s, 3H), 3.63 - 3.59 (m, 2H), 3.38 - 3.26 (m, 5H) 3.18 - 3.13 (m, 2H), 3.10 (s, 3H), 2.73 (t, J = 6.80 Hz, 2H), 2.07 - 2.02 (m, 1H), 1.35 (d, J = 9.60 Hz, 1H).

EXAMPLE S157.2 Synthesis of- ((3R, 4R) -4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidin-1-yl) -4- ((1-methyl-2-oxoindolin-5-yl) amino) pyrimidine-5-carbonitrile (157)

Step 1 Synthesis of 4- ((1-methyl-2-oxoindolin-5-yl) amino) -2- (methylthio) pyrimidine-5-carbonitrile. To a stirred solution of 5-amino-1-methylindolin-2-one (1.0 g,6.17 mmol) and 4-chloro-2- (methylthio) pyrimidine-5-carbonitrile (1.15 g,6.17 mmol) in THF (10 mL) was added DIPEA (2.7 mL,15.4 mmol). The reaction mixture was stirred at 25 ℃ for 16 h. The reaction mixture was filtered and the precipitated solid was washed with water (4×50 mL) to give the title compound as a yellow solid (1.77 g,5.58 mmol,91% yield).

Step 2 Synthesis of 4- ((1-methyl-2-oxoindolin-5-yl) amino) -2- (methylsulfonyl) pyrimidine-5-carbonitrile. To a stirred solution of 4- ((1-methyl-2-oxoindolin-5-yl) amino) -2- (methylthio) pyrimidine-5-carbonitrile (750 mg,2.41 mmol) in DCM (25 mL) was added mCPBA (1.25 g,7.23 mmol). The reaction mixture was stirred at 25 ℃ for 3h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (25 mL) and diluted with ethyl acetate (50 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as a yellow solid (715 mg,1.57 mmol,65% yield).

Step 3 Synthesis of 2- ((3R, 4R) -4- ((3- (2, 4-dioxotetrahydropyrimidin-1 (2H) -yl) -1-methyl-1H-indazol-6-yl) amino) -3-methoxypiperidin-1-yl) -4- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidine-5-carbonitrile. The title compound was synthesized according to general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (100 mg,0.26 mmol) and 4- ((1-methyl-2-oxoindolin-5-yl) amino) -2- (methylsulfonyl) pyrimidine-5-carbonitrile (103 mg,0.26 mmol) as starting material. LCMS [ M+H ] ⁺ found 636.2;¹H NMR (400 MHz, DMSO-d6): δ 10.46 (s, 1H), 9.29 (s, 1H), 8.40 (s, 1H), 7.48 - 7.43 (m, 2H), 7.30 (d, J = 8.80 Hz, 1H), 6.93 (d, J = 8.80 Hz, 1H), 6.57 (d, J = 8.80 Hz, 1H), 6.45 (s, 1H), 5.94 (s, 1H), 4.30 - 4.12 (m, 2H), 3.86 (t, J = 6.80 Hz, 2H), 3.81 (s, 3H), 3.65 (brs, 2H), 3.54 (s, 3H), 3.40 – 3.30 (m, 2H),3.25 – 3.12(m, 2H), 3.10 (s, 3H), 2.73 (t, J = 6.40 Hz, 2H), 2.08 (brs, 1H), 1.41 (brs, 1H).

EXAMPLE S158.Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4-fluoro-6- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (158)

Step 1 Synthesis of 5- ((5-chloro-2, 6-difluoropyrimidin-4-yl) amino) -1-methylindolin-2-one. To a stirred solution of 5-amino-1-methylindolin-2-one (500 mg,3.08 mmol) and 5-chloro-2, 4, 6-trifluoropyrimidine (519 mg,3.08 mmol) in ACN (25 mL) was added DIPEA (0.5 mL,3.08 mmol) at 0 ℃. The reaction mixture was stirred at 0 ℃ for 2 h. The reaction mixture was quenched with water (10 mL) and extracted with DCM (3×25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as an off-white solid (550 mg,1.59 mmol,52% yield).

Step 2 Synthesis of 1- (6- (((3R, 4R) -1- (5-chloro-4-fluoro-6- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione. The title compound was synthesized according to general procedure 6 using 1- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) dihydropyrimidine-2, 4 (1H, 3H) -dione (150 mg,0.39 mmol) and 5- ((5-chloro-2, 6-difluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (148 mg,0.43 mmol) as starting material. LCMS [ M+H ] ⁺ found 663.2;¹H NMR (400 MHz, DMSO-d6): δ 10.45 (s, 1H), 8.99 (s, 1H), 7.45 -7.49 (m, 2H), 7.29 (d, J = 8.80 Hz, 1H), 6.93 (d, J = 8.40 Hz, 1H), 6.56 (q, J = 1.60 Hz, 1H), 6.44 (d, J = 1.20 Hz, 1H), 5.91 (d, J = 8.00 Hz, 1H), 4.12 (d, J = 12.40 Hz, 1H), 3.86 (t, J = 6.80 Hz, 3H), 3.81 (s, 3H), 3.60 - 3.63 (m, 1H), 3.53 (s, 2H), 3.41 - 3.47 (m, 2H), 3.31 – 3.28 (m, 3H), 3.19 (s, 1H), 3.10 (s, 3H), 2.72 (t, J = 6.80 Hz, 2H), 2.01 - 2.07 (m, 1H), 1.50 - 1.60 (m, 1H).

Example S159.3- (6- (((3R, 4R) -1- (5-chloro-4-fluoro-6- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (159) synthesis

Step 1 Synthesis of 3- (6- (((3R, 4R) -1- (5-chloro-4-fluoro-6- ((1-methyl-2-oxoindol-5-yl) amino) pyrimidin-2-yl) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione. The title compound was synthesized according to general procedure 6 using 3- (6- (((3 r,4 r) -3-methoxypiperidin-4-yl) amino) -1-methyl-1H-indazol-3-yl) -3-methylpiperidine-2, 6-dione (150 mg,0.30 mmol) and 5- ((5-chloro-2, 6-difluoropyrimidin-4-yl) amino) -1-methylindolin-2-one (93 mg,0.30 mmol) as starting material. LCMS [ M+H ] ⁺ found 676.2;¹H NMR (400 MHz, DMSO-d6): δ 10.77 (s, 1H), 9.00 (s, 1H), 7.52 - 7.44 (m, 3H), 6.98 - 6.91 (m, 1H), 6.62 - 6.55 (m, 1H), 6.50 - 6.41 (m, 1H), 6.02 - 5.83 (m, 1H), 4.12 - 4.07 (m, 1H), 3.92 - 3.84 (m, 1H), 3.80 (s, 3H), 3.52 - 3.47 (m, 5H) 3.35 - 3.25 (m, 3H), 3.25 - 3.14 (m, 2H), 3.11 (s, 3H), 2.48 - 2.39 (m, 2H), 2.11 - 1.98 (m, 2H), 1.59 (s, 3H), 1.45 - 1.35 (m, 1H).

Formulation examples

Example f1 formulation of compound 70.

Compound 70 was selected for dosage form development. Compound 70 is a mixture of two diastereomeric compounds 71 and compound 72.

Film coated tablets of compound 70 were developed in two dosage forms, 20 mg and 80 mg.20 Tablets of the mg and 80 mg gauge contained a Spray Dried Dispersion (SDD) of compound 70 and inactive ingredients such as anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, sodium stearyl fumarate and Opadry film coatings. SDD contains 35% weight/weight (w/w) of compound 70, prepared by spray drying a solution of compound 70 and Eudragit L100-55 in methylene chloride and methanol. 20 The mg tablet was a powder gray round biconvex film coated tablet and the 80 mg tablet was a powder gray oval biconvex film coated tablet. Tablets are supplied in tightly closed High Density Polyethylene (HDPE) bottles with desiccant and child-resistant closure.

Biological examples

Example B1. BCL6_ HiBiT degradation assay.

DHL-4 cell lines were generated that knocked HiBiT-tags into the endogenous BCL6 locus using CRISPR. DHL-4_bcl6_hibit cells were dispensed into 384 well plates (Corning No. 3570) pre-spotted with compounds. Compounds were dispensed into 384 well plates by acoustic dispenser (ATS acoustic transfer system from EDC Biosystems) in a 10-point dose response curve (using 3-fold dilution starting from 1-10 μm and down to 0.0015-0.00015 μm). mu.L of medium (RPMI-1640+10% heat-inactivated FBS+1 XPen/Strep) containing 2500 DHL-4 was dispensed into each well. The assay plates were incubated at 37 ℃ with 5% CO ₂ for 2 hours. After incubation, 25 μl of Nano-Glo HiBiT lysis assay working solution (Promega, catalog No. N3040, madison, wisconsin) was added to each well and incubated 15-30 min at room temperature protected from light. After 30min, the luminescence was read on an Envision or PHERASTAR luminescence reader. To determine EC ₅₀ values (half maximum effective concentration) and gradient levels (Y constant) of BCL6 degradants, A four-parameter dose-response model in Dotmatics (y= (a+ ((B-A)/1+ ((C/x)) -D) was used to fit the luminescence signal (Y) normalized by DMSO control and compound concentration (x) ·the fitted value C was degradation EC ₅₀ and the minimum value A was the Y constant.

Degradation of BCL6 at 2h in DHL-4 results is shown in table 5, where EC ₅₀ is half maximal effect and Y _con is% of the remaining maximal effect.

Table 5.

* < 0.1 μM

* 0.1 To 1. Mu.M

*** > 1 μM

^ > 60%

^^ 20%-60%

^^^ < 20%

† Data from 24 h degradation assay

Example B2 phase I study of compound 70 alone and in combination with an anti-lymphoma agent in recurrent/refractory non-hodgkin's lymphoma (R/R NHL).

This study was a multicenter, open-label, dose discovery, phase 1 study to determine the safety, tolerability, PK, pharmacodynamics and preliminary efficacy of compound 70 in participants with R/R NHL. The study will be performed in two parts. Dose escalation (part a) compound 70 will be evaluated in participants with diffuse large B-cell lymphoma (DLBCL) and Follicular Lymphoma (FL) at increasing oral doses to evaluate safety, tolerability, and PK, and to guide determination of the Maximum Tolerated Dose (MTD) and/or Optimal Biologic Dose (OBD) of compound 70 as a single agent (part A1) or in combination with rituximab (part A2). Dose escalation (part B) will further evaluate the safety, primary efficacy and PK of compound 70 administered at or below MTD/OBD in one or more escalation cohorts to determine a recommended phase 2 dose (RP 2D) at one or more doses as a single agent (part B1) or in combination with an anti-lymphoma agent (part B2). Additional expansion queues evaluating other combinations (part B2) or selected patient populations (part B3) may be considered.

The participants will complete for up to three study periods, screening (up to 28 days [ +3 days ]), treatment (up to 104 weeks [ about 2 years ]) (from the first dose of study intervention, with or without treatment delay), and follow-up. The follow-up period included safety follow-up (28 days [ ±3 days after the last dose of study intervention ]), survival follow-up (up to 156 weeks [ about 3 years ] from the first dose of study intervention) and efficacy follow-up (up to 52 weeks [ about 1 year ]). The longest duration of study participation will be about 160 weeks (about 3 years) (up to 4 weeks of screening, up to 104 weeks of treatment, and up to 52 weeks of follow-up). Part A1 relates to BID administration of compound 70. Based on the emerging safety, PK and PD data, QD dosing exploratory cohorts can be opened and the starting dose will not exceed the dose shown to be tolerated in BID dosing.

A total of approximately 273 participants can be grouped, including 73 participants in part a (41 in part A1 and 32 in part A2), and 200 evaluable participants in part B (approximately 20 per queue: up to 80 in part B1 and B2, and up to 40 in part B3).

The inclusion criteria include:

participants must be over 18 years old or have R/R NHL (including DLBCL [ e.g., unspecific (NOS) DLBCL and diffuse large B-cell lymphoma/high grade B-cell lymphoma with MYC and BCL2 rearrangement ] and FL) according to legal adult age of local regulations, wherein one of the following requirements (as applicable to each indication) is recurrent or refractory disease:

For R/R DLBCL (de novo) and FL 3b, after at least two previous normals of therapy (e.g., having received a first line combination chemotherapy regimen containing rituximab, anthracyclines, alkylating agent, and steroid, and having received at least one additional [ second line/rescue ] treatment).

For R/R DLBCL (transformed lymphomas) after at least two prior normals to therapy (which must be administered after transformation; thus, at least two standard treatment regimens for DLBCL must have been accepted).

For R/R FL (except FL 3 b), the treatment criteria are met after at least two prior treatment norms and at the time of Group entry (e.g., according to but not limited to the follicular lymphoma study Group [ Group' Etude des Lymphomes Folliculaires, GELF ] criteria). The participants must have measurable disease, defined by having at least one Fluorodeoxyglucose (FDG) affinity lesion for FDG affinity disease and cross-sectional imaging by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI), having a two-dimensional measurable disease and at least one lesion transverse > 1.5 cm, as defined by the lukino classification of NHL (Lugano classification). Measurable disease cannot be previously irradiated.

Participants should have the following laboratory values:

absolute neutrophil count ≡1X10 ⁹/L and no growth factor support for 7 days (14 days in case of pefegrid)

-Aspartate Aminotransferase (AST) and alanine Aminotransferase (ALT) are 3.0. 3.0 x normal Upper Limit (ULN) or 5.0 x ULN if liver is affected

Total bilirubin is less than or equal to 1.5 x ULN

Potassium, magnesium and calcium (corrected Ca or free [ ionized ] Ca) concentrations are in the normal range, or can be corrected with supplements

Platelet count of ≡75 x 10 ⁹/L

Hemoglobin count of 9 g/dL or more

-Estimated serum creatinine clearance of ≡50 mL/min using kidney disease diet improvement (MDRD) method

Participants must receive and follow pregnancy prevention programs.

The exclusion criteria included:

Participants with physical stamina of equal to or greater than 3 in the eastern tumor cooperative group (ECOG).

Participants with non-hematologic toxicity (which does not regress, return to baseline or grade 2 or are considered irreversible) associated with previous or current anti-cancer therapies.

Participants with known dysphagia, short bowel syndrome, gastroparesis, or other conditions/procedures that limit digestion or gastrointestinal absorption of orally administered drugs.

Participants with uncontrolled hypertension (systolic BP > 180 mm Hg or diastolic BP > 100mm Hg) or hypotension (systolic BP < 90 mm Hg or diastolic BP < 60 mm Hg) at screening or within 1 month before C1D 1.

Clinically significant cardiovascular or cerebrovascular disease, including any of (i) complete left or double branch block or other clinically significant abnormal ECG findings at screening, (ii) history of congenital long QT syndrome, (iii) sustained or clinically significant ventricular arrhythmias at screening, (iv) QT interval corrected for heart rate using the friericia formula (QTcF) at screening for more than 470 milliseconds, (v) unstable angina, myocardial infarction or stroke or history of clinically significant arrhythmias (such as ventricular tachycardia, ventricular fibrillation or torsion of the tip) at less than 6 months prior to initiation of research intervention, (vi) heart association (New York Heart Association) with impaired cardiovascular function at more than 3 (level 3 is defined as heart disease resulting in significantly limited physical activity and participants feel comfortable at rest but less than normal levels of physical activity resulting in fatigue, palpitations, dyspnea or angina), LVEF < 45% at screening, as determined by ultrasound heart rate (ecga) or multiple gated (MUGA) scanning.

Treatment that is not subject to restrictions, precautions and contraindications.

If the participant is engaged in this study, the participant is placed at unacceptable risk for any condition (including significant acute or chronic medical conditions, active or uncontrolled infections, or the presence of laboratory abnormalities).

Participants with known SARS-CoV-2 infection with mild or asymptomatic infection within 10 days or severe/critical illness within 20 days before cycle 1, day 1.

A concurrent history of a second cancer requiring ongoing sustained systemic treatment.

Participants with a history of known or suspected central nervous system involvement or primary CNS lymphomas (including those in remission).

Previous concomitant therapy (i) previous live virus vaccine within less than or equal to 4 weeks before starting the intervention. (ii) The prior COVID-19 vaccine within 7 days prior to initiation of the intervention (for vaccines requiring more than 1 dose, when applicable and when delay in initiation of the study intervention does not put study participants at risk, then the full series (e.g., two doses of the 2 dose series) should be completed at least 7 days prior to initiation of the study intervention, (iii) the participants have received prior therapy of ∈5 half-life or have received prior systemic anti-cancer therapy (approved or experimental) 4 weeks prior to initiation of the study intervention (on the basis of the shorter), (iv) the participants have received prior CAR-T or other T cell targeting therapy (approved or experimental) 4 weeks prior to initiation of the study intervention, (v) the participants have received prior therapy with a drug that modulates Cerebulin (CRBN) 4 weeks prior to initiation of the study intervention (e.g., lenalidomide, atorvastatin (avadomide), pomalidomide, CC-99282), (vi) the participants are using chronic systemic immunosuppressant therapy or corticosteroid (e.g., approved or experimental) and (in vitro) the clinical conditions allowing the transient to be inhaled by the human eye beyond the hd of mg or by the local human eye beyond the allowable limits of the clinical criteria of the hd (hd) and the clinical conditions.

Participants received prior autologous SCT within less than or equal to 3 months prior to the initiation of study intervention. If the participants received prior autologous SCT within > 3 months prior to the beginning of the study intervention, then any treatment-related toxicity did not resolve (> grade 2).

Participants received standard or reduced strength pretreatment of prior allogeneic SCT within less than or equal to 6 months prior to initiation of study intervention. If the participants received prior allogeneic SCT within > 6 months prior to the beginning of the study intervention, then any treatment-related toxicities did not resolve (> grade 2).

Participants received major surgery less than or equal to 2 weeks before the study intervention was initiated. The participants must have recovered from any clinically significant effects of recent surgery.

There is evidence for the presence of organ dysfunction or any clinically significant deviation from normal (which is outside of the range consistent with the target population) in physical examination, vital signs, ECG or clinical laboratory determinations.

Participants with known HIV seropositive or active infection, or active infection with Hepatitis B Virus (HBV) or hepatitis C virus (as evidenced by positive screening of blood for hepatitis C antibodies; hepatitis B surface antigen and anti-hepatitis B surface antibodies, or HIV-1 and HIV-2 antibodies).

History of allergy to rituximab or any excipient.

Compound 70 will be administered orally continuously twice daily (BID) over a 28 day period starting on cycle 1 day 1 until clinically significant disease progression, unacceptable toxicity, or participant/doctor decision to exit for up to 2 years of total treatment. Alternative schedules include a 21/28 day schedule and a 14/28 day schedule, or once daily (QD) in any of these schedules. Rituximab will be administered at a fixed dose of 375 mg/m ² for up to six infusions (days 1-6 cycles 1) according to the drug instructions and institutional standard specifications. Survival follow-up lasted up to 3 years (156 weeks) from the first treatment. Participants can enter the group study for up to about 160 weeks (including screening, treatment, and follow-up periods).

For part A1, a Bayesian Optimization Interval (BOIN) design for phase 1 will be followed by a TITE-BOIN design for phase 2 to guide the making of the dose escalation decision and the determination of the MTD and/or the OBD(s). For part A2, the dose up/down decision will be guided by BOIN. For part B, the participants will be randomly assigned to an extended cohort of approximately 20 evaluable participants per dose level/schedule, each cohort receiving compound 70 alone or in combination with an anti-lymphoma agent. Will exhibit a dose-level DLT rate and frequency distribution of participants with Adverse Events (AEs). Summarized statistics of PK parameters will be provided. The point estimate of ORR generated by the krobot-Pearson method (Clopper-Pearson method) was combined with a 95% Confidence Interval (CI). The median time to event endpoint (e.g., progression Free Survival (PFS)) and 95% CI will be estimated using KM methods.

Incremental doses of compound 70 will be evaluated in part a. The temporary dosing cohorts for compound 70 dose escalation are summarized in table 6. The initial dose of compound 70 in part A1 will be 40mg for BID oral administration. If the initial dose of compound 70 is intolerant, lower dose levels (e.g., cohort-1, 20 mg BID) can be explored. The temporary dose increments will follow a modified fibonacci design from 80 mg BID and above, where doses in successive dose cohorts can be increased by 100%, 62.5%, 53.8%, 33%, etc. of the doses in the previous cohorts. Smaller increments may be recommended for dose escalation when considering all available clinical data. Based on emerging safety, PK and pharmacodynamic data, QD dosing exploratory cohorts can be opened and the starting dose will not exceed the total daily dose shown to be tolerated in BID dosing. Alternative dosages, schedules and frequencies may be proposed. In all cases, the recommended dose for the next cohort will not increase by more than about 100% relative to the previous total daily dose.

Table 6.

The starting dose and schedule of compound 70 in part A2 will be determined based on the emerging safety, PK and pharmacodynamic data from parts A1 and B1, and will be at least one dose level lower than the maximum compound 70 monotherapy dose explored in part B1. The initial dose of compound 70 in part A2 will be determined using data from a minimum of 25 participants in monotherapy treated with OBD.

After the dose escalation is completed, the expansion may be performed in compound 70 OBD and/or alternative tolerable dosing schedules in a plurality of queues allocated in a randomized fashion. The decision to dose extend at a given dose level and schedule will be based on, but not limited to, the overall safety profile, preliminary positive efficacy signal, and available PK/pharmacodynamic data. Approximately 20 participants at each dose level/schedule and with sufficient follow-up will enter the B1 and B2 fractions to further evaluate the safety and efficacy of compound 70 alone or in combination with rituximab administered at OBD and/or lower dose levels and/or alternative schedules. Two dosage schedules will be evaluated for each indication (part B1) or each combination (part B2). The data of participants treated at OBD and/or lower dose levels and/or alternative schedules in dose escalation may be combined with the data of participants treated at the same dose levels, schedules and treatment arms in dose escalation (if from the same population). Additional expansion queues that evaluate other combinations or selected patient populations may be considered and will be included in part B2 or part B3 via a protocol revision. Toxicity will be monitored in monotherapy and combination expansion cohorts using bayesian continuous monitoring.

Compound 70 will be administered BID continuously orally in a 28 day cycle starting on cycle 1, day 1 (C1D 1). Alternative schedules include a 21/28 day schedule (21 day dosing/7 day dosing) and a 14/28 day schedule (14 day dosing/14 day dosing) over a 28 day period, or a once daily (QD) dosing in any of these schedules. Participants receiving BID administration must take tablets of compound 70 at about the same time each day in the morning and evening. The interval between 2 doses should be about 12 hours ± 2 hours. Participants receiving QD dosing will take compound 70 once daily in the morning at about the same time.

Participants should avoid eating Seville orange (Seville orange), grapefruit or grapefruit juice [ grapefruit, exotic citrus fruits, grapefruit hybrids or fruit juice ] 5 days before the start of study intervention until 24 hours after the final dose of compound 70.

Participants in all groups were asked to fasted (not to eat or drink anything other than water) about 2 hours before administration of compound 70 until 1 hour after administration. In some instances, participants may be allowed to avoid food and beverage (other than water) restrictions 2 hours before and 1 hour after administration of compound 70. If the participants miss a dose of compound 70, the dose will still be available for administration within up to 6 hours for BID administration (or up to 12 hours for QD administration) after the time they would normally take the dose. If more than 6 hours (or 12 hours for QD dosing) pass, they should be instructed to skip the missing dose. At the next scheduled dose, they should take compound 70 at the usual time. Participants should not take 2 doses to compensate for their missing dose.

If the participant vomits within 30 minutes of ingestion of compound 70, the compound 70 tablet may be ingested again. If emesis occurs after 30 minutes of compound 70, the participants must not ingest additional tablets of compound 70 unless the entire drug is visible in the vomit.

Compound 70 may be administered for up to 104 weeks (about 2 years) or until clinically significant disease progression, unacceptable toxicity, or withdrawal of consent. Retreatment with compound 70 may be considered if (i) the participant discontinued treatment for reasons other than AE, progression, or initiation of new anti-lymphoma therapy, (ii) the participant achieved a response demonstrated with compound 70 based on malignant lymphoma international working group criteria (The International Workshop Criteria for Malignant Lymphoma), (iii) the participant had evidence of disease recurrence based on malignant lymphoma international working group criteria during the lifetime follow-up period, (iv) continued qualification. Participants receiving retreatment will follow the schedule of activity starting on day 1 of cycle 1, but may continue with their most recent dose level and a schedule or dose level at which at least one participant cohort in the study shows adequate tolerance. The participants were treated only once more during the study.

Rituximab will be administered at a dose of 375 mg/m ² at day 1 of cycles 1 to 6 as an IV infusion. Dose reduction of rituximab is not allowed. Rituximab may be administered with a morning dose of compound 70 or within up to 2 hours after a morning dose of compound 70.

Suggested body surface area estimation methods include Mosteller and DuBois.

Bsa= v ((height (cm) x weight (kg))/3600

Bsa= 0.20247 x height (m) 0.725 x weight (kg) 0.425

Prior to each rituximab infusion, a precursor drug consisting of acetaminophen and an antihistamine should be administered (see current local rituximab prescription information). Depending on institutional practices, steroids may also be administered prior to the onset of rituximab infusion. Monitoring measures during and after infusion of rituximab should be applied as recommended by the manufacturer and current guidelines.

Rituximab may be administered in an in-patient or out-patient clinical setting. For dose adjustment, recommendations follow the drug specifications and local practices. Dose reduction of rituximab is not allowed.

Participants who do not require dose interruption are allowed to enter the next treatment cycle (e.g., hematologic toxicity must resolve to < 2). If rituximab doses are discontinued due to study intervention-related toxicity, rituximab may be restarted once toxicity subsides to < grade 2. In cycle 1, the skipped dose due to toxicity will not be rescheduled. In a period of ≡2, the missing dose of rituximab can be made up within 2 weeks (i.e. days 1 to 14 of the period). If toxicity subsides to < 2 orders during days 15 to 28 of the cycle, rituximab dosing will resume on day 1 of the next cycle. Participants will be closely monitored throughout the infusion period and for at least 1 hour after continuous infusion, and a series of vital signs are obtained and pulse oximetry monitoring is performed. In participants with grade 4 infusion reactions or recurrent grade 3 infusion reactions requiring permanent rituximab discontinuation, additional blood tests can be obtained within 15 to 60 minutes of the onset of symptoms to assess severe allergic reactions. Laboratory tests may include immunoglobulin E (IgE), total tryptase, and plasma histamine.

In a phase 1 study evaluating compound 70, a series of pharmacodynamic, predictive, and exploratory biomarkers will be collected to facilitate dose/schedule selection, and potentially identify biomarkers to predict response and inform potential combinations.

The primary pharmacodynamic biomarker for compound 70 is expected to be degradation of BCL6 protein. For this purpose, changes in BCL6 level expression from baseline will be measured in Peripheral Blood Mononuclear Cells (PBMCs). Samples collected via Fine Needle Aspirate (FNA) or tumor biopsy were used to measure BCL6 and downstream markers using flow cytometry-based assays to identify potential markers of target engagement and response to compound 70 in tumor cells. Longitudinal quantification of apoptosis of circulating tumor deoxyribonucleic acid (ctDNA) and Lymph Node (LN) tumor cells was performed as a means to assess early effects of study treatments on overall tumor burden. To explore the evolving immune response and dose schedule effects of compound 70 as a single agent and its combination with an anti-lymphoma agent, immune-related assessment would include analysis of immune profiles of T cell, B cell and NK cell subsets and cytokine secretion to be assessed prior to and during treatment with compound 70. Predictive markers of response to single agent compound 70 and its combination with other anti-lymphoma agents will be explored at baseline as well as in the study treatment being conducted. These studies will include collection of paired tumor biopsies before and during treatment. Assays based on RNA profiling, BCL6 protein expression levels, and immune cell infiltration (e.g., T cells, B cells, macrophages, and/or NK cells) characterization would be explored as potential predictors of response. The diagnostic biopsy will also be analyzed for gene expression characteristics that may be associated with enrichment responses to compound 70 as a single agent or in combination with other anti-lymphoma agents, including downstream markers (e.g., c-Myc, NCOR1/2, PELI 1) and biomarkers associated with poor prognosis (e.g., COO, c-Myc, BCL2 or BCL6 gene rearrangements, c-Myc, BCL2 or BCL6 protein expression, mutation status of lymphoma-associated genes, etc.). Biomarker assessment will be performed according to the laboratory manual.

Table 7 lists the sample types used for the various biomarker evaluations of the plans and the analytical techniques of the plans.

Table 7.

Example B3.

This example is incorporated herein by reference in its entirety for the disclosure of the researcher manual (investor's Brochure) (version number: 01; version date: 2023, month 6, 21).

References to chapters, tables, references, and/or appendices in example B3 refer to other chapters, tables, references, and/or appendices in example B3.

List of abbreviations

ADME absorption, distribution, metabolism, and excretion

AE adverse events

ALT alanine aminotransferase

ANOVA analysis of variance

Area under AUC concentration-time curve

Area under the concentration-time curve for AUC (INF) from 0 to infinity

BCL 6B cell lymphoma 6

BCRP (BCRP) breast cancer drug-resistant protein

BID twice a day

BLQ is below the limit of quantitation

BMSBristol-Myers Squibb

BSA body surface area

BSEP bile salt output pump

BUN blood urea nitrogen

Caco-2 high-pressure sorrel intestinal adenocarcinoma cell line

Cmax maximum observed concentration

Xenografts derived from CDX cell lines

CELMoD cerebellar protein E3 ligase modulating medicine

CLTp plasma clearance

COO origin cell

Coefficient of variation of CV

CYP human cytochrome P450

DBP diastolic pressure

Half maximum degradation concentration of 50% protein degradation by DC50

DCN file control number

DDI drug-drug interactions

Diffuse large B-cell lymphoma of DLBCL

DMAC dimethylacetamide

ECG electrocardiogram

EMA European drug administration

F absolute oral bioavailability

FDA U.S. food and drug administration

FIH first time human body

FL follicular lymphoma

FOB fecal occult blood

Non-binding fraction of fu

Unbound fraction in fu, p plasma

GALT intestinal associated lymphoid tissue

GC hair-growing center

GLP good laboratory Specification

HDPE high density polyethylene

HEK human embryo kidney

HiPSC-CM human induced pluripotent stem cell-derived cardiomyocytes

HPLC high performance liquid chromatography

HNSTD highest non-severe toxic dose

HPMC-AS-HF hydroxypropyl methylcellulose acetate succinate

Concentration at which 50% inhibition was observed for IC ₅₀

ICH International conference of coordination

IFN interferon

IL interleukins

IV intravenous

Ka absorption Rate constant

KI inhibition constant

Kinact enzyme inactivation Rate

KLH keyhole limpet hemocyanin

LC-MS/MS liquid chromatography tandem mass spectrometry

LC-UV/MS liquid chromatography-ultraviolet spectroscopy/mass spectrometry

LDD ligand directional degradation agent

LLOQ quantitative lower limit

MAP mean arterial pressure

MATE multidrug and toxin efflux proteins

MDCKMADIN-Darby canine kidney epithelial cells

MDR1 multidrug resistance 1 gene

MRNA

MRP multi-drug resistance related protein

Maximum recommended initial dose of MRSD

Mean residence time of MRT

Maximum tolerated dose of MTD

NA not available

NADPH nicotinamide adenine dinucleotide phosphate

NHL non-Hodgkin lymphoma

NK natural killer cells

Level of no adverse effects observed by NOAEL

OATP organic anion transport polypeptides

OCT organic cation transporter

OECD economic Cooperation and development organization

P-gpP-glycoprotein

PD pharmacodynamics

Xenografts of PDX patient origin

PEG-400 polyethylene glycol 400

PK pharmacokinetics

PMBL primary mediastinal B cell lymphoma

PO oral administration

QC quality control

RBC red blood cells

R-CHOP rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone

R/R recurrent/refractory

SBP systolic blood pressure

SDD spray-dried dispersions

Sf9 spodoptera frugiperda 9

Severe toxic dose of 10% or less in STD10 animals

Apparent terminal HALF-life of T-HALF

Tfh helper T cells

Time to maximum observed concentration of Tmax

THL/TEL triple-hit lymphoma or triple-expression lymphoma

TK toxicology

TNF tumor necrosis factor

UDP uridine diphosphate

ULOQ upper limit of quantitation

UGT glucuronyl transferase

US united states

UV ultraviolet light

Vss steady state distribution volume

WBC white blood cells

1. Summary. This investigator manual contains information about BMS-986458, a small molecule bifunctional cerebulin-dependent Ligand Directed Degradant (LDD) of BCL6, originally developed for treating patients with recurrent and/or refractory diffuse large B-cell lymphomas (R/R DLBCL) and follicular lymphomas (R/R FL). DLBCL is an aggressive subtype of non-hodgkin lymphoma (NHL), accounting for approximately 4% of all cancers in the United States (US) as a whole, with an estimated 29,108 new cases ¹ in the united states in 2020 accounting for 26% of all cases of diagnosed NHL neoplasms. R/R FL is the second most common lymphoma diagnosed in the United states and Western Europe, accounting for approximately 22% of NHL cases, and 70% are indolent lymphomas. ²

BMS-986458 is a mixture of two diastereomers. Two of the three stereogenic centers in each isomer have the (R) configuration. The third stereogenic center has the (R) configuration in BMT-648592 and the (S) configuration in BMT-648599. Individual isomers were also tested in pharmacokinetic and pharmacodynamic studies. The individual diastereomers showed equivalent activity ³ in all in vitro pharmacological tests performed. In vivo OCI-LY-1 xenograft models, similar degrees of tumor BCL6 degradation ¹⁵ were observed for BMS-986458 and its individual diastereomers at the same dose level. Additional chiral analysis showed that significant epimerization occurred in mice.

Pharmacological, safety pharmacological and Pharmacokinetic (PK) evaluations have been performed on BMS-986458. A critical well-defined laboratory specification (GLP) toxicity study was performed in rats and dogs using the oral route (the intended route of administration in clinical trials) to assess and characterize the potential toxicity of BMS-986458.

1.1 Background. The current standard of care for DLBCL is an intensive immunochemical regimen consisting of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP). Addition of monoclonal anti-CD 20 rituximab to CHOP therapy resulted in a 15% increase in total survival, but despite this improvement, 40% of patients eventually experience recurrent or refractory disease and represent a large area of unmet medical need. ^4,5,6,7,8,9

Clinical studies have demonstrated a number of cytogenetic and mutation profiles associated with high risk NHL, with the most damaging being defined by perturbations involving triplets of oncogenes c-Myc, BCL2 and BCL6, termed triple-hit lymphoma or triple-expression lymphoma (THL/TEL). In THL/TEL, these factors are either genetically translocated within the immunoglobulin locus, resulting in exogenous upregulation, or exhibit a mutant character with regulatory factors driving oncogene overexpression or protein stabilization. Together, these oncogenes lead to abnormal proliferation of transformed B-cell lymphomas and resistance to therapeutic agents.

BCL6 is the primary transcriptional regulator that controls B cell differentiation and antibody affinity maturation programs. BCL6 expression in activated B cells is elevated upon transition into Germinal Centers (GC) and promotes tolerance to genotoxic stress associated with somatic hypermutation and class switch recombination (the process required to differentiate into plasma or memory B cells expressing or secreting high affinity antibody B cell receptors). In 20% of newly diagnosed DLBCL patients expressing exogenous levels of BCL6, cell cycle checkpoints and DNA damage response pathways were pleiotropic inhibited, resulting in unregulated cell proliferation and malignant transformation. Degradation and genetic perturbation of BCL6 in DLBCL cell line models has shown that upon de-inhibition of downstream processes, the cell cycle is prevented or compromised and DNA damage response is activated, ultimately leading to apoptosis and cell death.

BMS-986458 potently and selectively degrades BCL6 in both the DLBCL cell line and normal immune cells. In vitro, BMS-986458 induced acute apoptotic responses in a subset of DLBCL cell lines expressing high levels of BCL6 protein, which was ubiquitous in the center of development B cell (GCB) cell-derived (COO) model. In all cases, both apoptosis and antiproliferative response to BMS-986458 were significantly more potent than the cerebellar protein E3 ligase modulating drug (CELMoD) lenalidomide. Enhanced sensitivity to BMS-986458 was observed in the R/R DLBCL model relative to the control group. In addition to in vitro activity, BMS-986458 also exhibited strong in vivo anti-tumor activity in two DLBCL cell line-derived xenograft (CDX) models and anti-proliferative responses from ex vivo patient-derived xenograft (PDX) models.

1.2 Non-clinical pharmacology. BMS-986458 was evaluated in several in vitro NHL models and showed rapid and potent antiproliferative and pro-apoptotic tumor cell killing in the broad spectrum GCB and primary mediastinal B cell lymphoma (PMBL) subtypes of DLBCL. The anti-tumor response was also evaluated in several models of acquired resistance to the standard care agent doxorubicin, which exhibited significant sensitization to BMS-986458 and a shift from anti-proliferative to apoptotic response relative to doxorubicin-sensitive controls.

Broadly, the anti-tumor response to BMS-986458 is associated with BCL6 expression levels and thus with GCB subtype patterns enriched with perturbations that lead to BCL6 upregulation or protein stabilization. The nature of the response from BCL6 expressing cell line models is subdivided into two categories, acute apoptosis or delayed anti-proliferative response. The acute response was shown to be apoptotic signaling within 5 days of compound treatment, whereas the late sensitizer showed a negligible or mild effect on proliferation after 5 days, which was then enhanced after BMS-986458 further exposure for 5 days. Cell line models that do not express BCL6 do not exhibit any loss of proliferative capacity after prolonged compound exposure.

The mechanism of action of BMS-986458 requires interaction with the CRL4CRBN E3 ubiquitin ligase substrate receptor cerebellar protein (CRBN) to induce recruitment of the target protein BCL 6. Subsequent BCL6 ubiquitination triggers proteasome destruction of the target, leading to growth inhibition and induction of apoptosis. Furthermore, proteomics and mechanism studies demonstrated that BCL6 is the only substrate degraded by BMS-986458 in the DLBCL model evaluated, and that modulation of several downstream pathways mediate the antiproliferative activity of BMS-986458. Within 24 h of the drug treatment, key mediators and antiproliferative proteins are de-inhibited, including cell cycle checkpoint inhibitors p27/CDKN1 and PTEN/SHP1 (a key inhibitory phosphatase).

In vivo, the antitumor activity of BMS-986458 was evaluated in the OCI-LY-1 and WSU-DLCL2 DLBCL xenograft models (representing preclinical models of acute and delayed sensitivity, respectively) with a twice-a-day (BID) dosing regimen. Initial pharmacokinetic/pharmacodynamic (PK/PD) studies in tumor-bearing mice established a clear correlation between tumor concentration of BMS-986458 and depth/duration of BCL6 degradation in tumor tissue. BID dosing regimen PK/PD studies resulted in plasma and tumor concentrations suitable for inducing near complete degradation (95% to 100%) of BCL6 in tumors within 24 h. In both DLBCL xenograft models, BMS-986458 exhibited antitumor activity in vivo in BID dosing mode, achieving significant Tumor Volume Reduction (TVR). This TVR correlates with PK exposure and depth/duration of BCL6 degradation. PK and PD analysis of tumors collected at the end of these studies revealed significant plasma and tumor concentrations of BMS-986458, and sustained degradation of ≡95% of tumor BCL6 appears to be necessary to show a minimum of about 65% tumor volume reduction in OCI-LY-1 xenograft models. In the OCI-LY-1 xenograft model, this degree of tumor BCL6 degradation and corresponding anti-tumor response can be achieved with repeated BID dosing of 15-30 mg/kg BMS-986458. In vivo OCI-LY-1 xenograft models, a similar degree of tumor BCL6 degradation was observed for BMS-986458 and its individual diastereomers at the same dose level. Furthermore, at the same dose level, similar responses were observed in the in vivo WSU-DLCL-2 xenograft model. Additional chiral analysis indicated that significant epimerization occurred in mice.

Within GC, B cells circulate between dark and bright areas, as they undergo successive rounds of affinity maturation. In the open region, B cells interact with follicular helper T cells (Tfh), where they facilitate the selection and survival of high affinity clones through cell-cell interactions and cytokine signaling. Tfh cells are also required to maintain GC structure, and therefore are critical for the production of high affinity antibodies upon immune attack. Survival of a subtype of NHL that proliferates within the lymph nodes, such as Follicular Lymphoma (FL), depends on Tfh support. BCL6 is a lineage determinant of Tfh cells, which is required to maintain cell identity and function within GC. The effect of BMS-986458 on the induced population of Tfh was evaluated in vitro, where it was shown that degradation of BCL6 resulted in loss of permeability of Tfh cell identity through expression of canonical markers CXCR5 and PD 1.

Although the molecular pathway downstream of BCL6 degradation is critical for lymphoid differentiation and function, there may be potential biological effects in other immune populations as well. Neutropenia is a common class II AE observed in many current hematological therapeutics, and thus the effect of BMS-986458 treatment on neutrophil viability and maturation pathways was evaluated in vitro. No loss of viability of cd34+cd33-CD11 b-cells matured to the CD34-CD33-cd11b+ stage was observed at clinically relevant concentrations compared to vehicle controls.

1.3 Non-clinical pharmacokinetics. In vitro and in vivo studies have been performed to characterize the absorption, PK, distribution, metabolism and drug interaction potential of BMS-986458.

BMS-986458 showed low to moderate systemic clearance (≤44% of liver blood flow), low to moderate steady state distribution volume (Vss; 0.4-3.5 times the total systemic water), moderate oral bioavailability (22% to 37%) in mice, rats, dogs and monkeys, with a corresponding apparent terminal half-life ranging from 0.9 to 5.5 hours. Furthermore, significant interconversions (up to 41% based on AUC) between diastereomers of BMS-986458 (BMT-648592 or BMT-648599) were observed in rats, dogs and monkeys after IV infusion or PO administration of the individual diastereomers.

BMS-986458 was highly bound to plasma proteins in mice, rats, rabbits, dogs, monkeys, and humans, with an unbound fraction (fu) < 1%. Species differences of fu were observed between humans and dogs (5.3×), whereas no significant differences were observed between humans compared to rats or rabbits (2×). Brain BMS-986458 concentration was < 0.22. Mu.M after a single oral dose of 10 to 300 mg/kg to Sprague-Dawley rats (brain/plasma ratio was ≡0.10 at ≡100 mg/kg and not properly assessed at ≡30 mg/kg, as concentrations were low or not measurable in female and male rats at 24 and 48 hours post-dosing, respectively), indicating minimal penetration across the blood brain barrier.

Following oral administration, metabolism of BMS-986458 was evaluated in human hepatocytes, as well as hepatocytes from preclinical species, and in plasma samples from rats and dogs. The primary metabolic pathways of BMS-986458 in the species tested include demethylation, oxidation, followed by glucuronidation and hydrolysis. BMS-986458 is the primary drug-related circulating compound in rat and dog plasma. Although there were quantitative differences in metabolite formation between species, qualitatively, all metabolites formed in human hepatocytes were also formed in hepatocytes from rats and/or dogs (both species for preclinical safety testing), indicating the absence of unique human metabolites.

Reactive phenotyping studies of the human cytochrome P450 (CYP) subtype and human liver microsomes indicate that BMS-986458 is metabolized primarily by CYP3A4/5 and that CYP1A2, CYP2C8, CYP2C9 and CYP2C19 are less involved. In vitro, BMS-986458 exhibited time-dependent inhibition of CYP3A4, with an enzyme inactivation rate (kinact) of 0.054/min and an inhibition constant (KI) of 28. Mu.M. In human hepatocytes, BMS-986458 up to 20. Mu.M showed weak induction of CYP1A2 and no induction of CYP3A4 or CYP2B 6. These data indicate that there is potential for drug-drug interactions (DDI) if BMS-986458 is co-administered with strong inhibitors or inducers of CYP3A4, sensitive CYP3A substrates, or CYP3A substrates with narrow therapeutic windows.

In addition, BMS-986458 showed inhibition of Uridine Diphosphate (UDP) -glucuronyl transferase (UGT) 1A1 with an IC50 of 0.15. Mu.M.

In vitro assays indicate that BMS-986458 is a substrate for the efflux transporter P-glycoprotein (P-gp) and most likely for the Breast Cancer Resistance Protein (BCRP). BMS-986458 is also an inhibitor of P-gp and BCRP, with half maximal inhibitory concentrations (IC 50) values of 3.3 and 1.6. Mu.M, respectively. These data indicate that BMS-986458 has the potential to cause inhibition of P-gp and/or BCRP in the gut and that absorption of BMS-986458 may be increased in the presence of P-gp inhibitors.

Furthermore, BMS-986458 showed inhibition of various liver and gall and kidney transporters by Organic Anion Transport Polypeptide (OATP) 1B1, OATP1B3, multidrug resistance-associated protein (MRP) 2, organic Cation Transporter (OCT) 1, OCT2, bile Salt Efflux Pump (BSEP), multidrug and toxin efflux proteins (MATE) 1 and MATE2K, wherein IC50 values range from 0.29 to 9.5. Mu.M. Overall, the potential for systemic interactions with these drug transporters appears to be low based on the expected clinical exposure (within the expected effective dose range). However, since clinical exposure is available, the potential for DDI will be continuously assessed.

1.4 Non-clinical toxicology. The non-clinical toxicology of BMS-986458 (a mixture of diastereomers BMT-648592 and BMT-648599) was evaluated in a series of comprehensive non-clinical toxicity studies including single and repeat dose toxicity studies for BID administration in rats and dogs for a duration of up to 28 days, in vitro and in vivo genotoxicity studies, in vitro phototoxicity studies, and in vitro and in vivo cardiovascular safety pharmacological studies. The exposure fold shown below was calculated from the predicted exposure (AUC and Cmax) at the proposed 40 mg BID initial clinical dose. The Cmax fold has been corrected for free fraction using a flux method using predicted species differences in protein binding and by assuming 99% protein binding for in vitro assays.

The potential of BMS-986458 to affect the cardiovascular system has been characterized both in vitro and in vivo. BMS-986458, BMT-648592, and BMT-648599 inhibited cardiac ion channels in vitro at approximately comparable concentrations, with hERG IC50 of 0.44, 0.52, and 0.16 μg/mL (0.7, 0.83, and 0.25 μM), respectively (519× for BMS-986458). BMT-648592 and BMT-648599 inhibited cardiac L-type calcium channels by 9.4% and 7.4%, respectively, at the highest test concentration of 0.628 μg/mL (1 μM). In a multi-electrode assay in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM), BMS-986458 increased field potential duration (FPDc), arrhythmia beats and decreased conduction velocity at a concentration of ≡0.19 μg/mL (0.3 μM;224 x), and an increase in beat frequency was observed at 1.9 μg/mL (3 μM;2240×). In exploratory rabbit electrophysiology studies, an increase in QTc interval (friericia corrected QT [ QTcF ] = 51.2 ms;Van de Water corrected QT [ QTcV ] =24.4 ms) was noted when 12 mg/kg BMS-986458 (free plasma concentration=1.16 μg/mL,0.729 μm;1365×) was administered by IV infusion, but an increase in heart rate (34%) and a decrease in mean blood pressure (-34%) interfered with the explanation. No BMS-986458 related cardiac, neurological and/or respiratory findings were noted in rats (free Cmax=0.194 μg/mL;228×) at doses up to 500 mg/kg/BID and in dogs (free Cmax=0.866 μg/mL;192×) at doses up to 50 mg/kg/BID in non-GLP studies telemetry rats and dogs and in 1 month GLP rats and dogs studies.

BMS-986458, BMT-648592 and BMT-648599 were mutagenic negative in Ames assays in vitro, but positive for fragmentation (clastogenicity) and aneuploidy (aneugenicity) in the presence or absence of S9 component in exploratory in vitro assays in CHO and/or TK6 cells. BMS-986458-related effects measured on peripheral micronuclei or comets (duodenum and liver) were not noted at daily doses up to 500 mg/kg BID in non-GLP 2-week rat toxicity studies and in independent GLP-compliant in vivo genotoxicity studies, respectively, indicating that BMS-986458 is genotoxic in vivo.

In the neutral red uptake phototoxicity assay consistent with GLP in Balb/c 3T3 mouse fibroblasts, BMS-986458 was positive for phototoxicity potential. Follow-up confirmation of phototoxic potential will be performed in vivo studies.

Repeated dosing toxicity studies were performed in rats and dogs for a duration of up to 1 month. In key 1 month oral toxicity studies (doses of 50, 150 and 500 mg/kg BID) performed in rats, no adverse findings were present at high doses up to 500 mg/kg BID (1000 mg/kg/day total). Thus, 500 mg/kg BID was considered as a level at which no adverse effect was observed (NOAEL). BMS-986458 was orally administered at doses of 0, 5, 15 and 50 mg/kg BID in a critical 1 month oral toxicity study in dogs. At a dose of 50 mg/kg BID, a decrease in food consumption was noted on day 1, leading to weight loss and a decrease in clinical symptoms, requiring early euthanasia of 3 out of 10 dogs on day 7, and no clear cause of clinical state decline (CLINICAL DECLINE). All other dogs at 50 mg/kg BID survived to scheduled necropsy without significant clinical signs. No other adverse findings were present and BMS-986458 was well tolerated at doses of 15≤ 15 mg/kg BID. Thus, NOAEL and the highest non-severe toxic dose (HNSTD) were considered to be 15 mg/kg BID.

In the repeated dose toxicity study, several dose-responsive non-adverse findings were observed in rats and dogs. Consistent with the role of BCL6 as an immune system modulator, ^10,11 immune suppression and pro-inflammatory changes were observed in rats and dogs, including increases in circulating WBCs, neutrophils, lymphocytes and T cells, B cells and/or Natural Killer (NK) cells, decreased production of anti-Keyhole Limpet Hemocyanin (KLH) IgM and/or IgG antibodies, decreased cytokines (interleukins [ IL ] -1 beta, IL-2, IL6, tumor necrosis factor [ TNF ] alpha, interferon [ IFN ] gamma) in rats alone, decreased amounts of lymphoid cells in thymus, gut-related lymphoid tissue (GALT) and lymph nodes (dogs) or spleen border areas (rats), mixed cell inflammation of femoral tibial joints in 4 dogs, and minimal neutrophil inflammation of atrioventricular valves in 1 dog (both findings were only at high doses). These findings are consistent with published roles of BCL6 as a major regulator of the immune system. A decrease in Red Blood Cell (RBC) parameters with regeneration reactions was observed in both species. In rats, an increase in platelets was observed, with no consistent change in coagulation parameters in each study. An increase in bilirubin was noted in dogs, with no microscopic related abnormalities (correlate) or transaminase increase, and potentially mediated by UGT1A1 inhibition. Liver hypertrophy and weight gain were observed in both species, with findings observed in rats associated with enzyme induction and secondary thyroid epithelial hypertrophy. In dogs, inconsistent changes in stool composition and positive Fecal Occult Blood (FOB) tests were observed without associated gastrointestinal pathology. Less kidney effect and no pathology-related abnormalities were noted in both species, with an increase in kidney weight in dogs and a slight increase in Blood Urea Nitrogen (BUN) in rats. Finally, minimal hypertrophy of the fascicular bands was observed in rats.

BMS-986458 does not degrade classical CELMoD new substrates such as Aiolos, CK1 alpha, ikaros, GSPT1 and SALL4. Thus, it is expected that BMS-986458 toxicity profile will differ from CELMoD, including potential reproductive toxicity. However, since reproductive toxicity assessment has not been performed so far, and the mechanism of BMS-986458 is cerebellar protein-dependent, BMS-986458 was treated as a putative selective developmental toxic substance.

Overall, non-clinical toxicology assessment supports the progression of BMS-986458 to clinical trials.

1.5 First time human clinical study. Study CA123-1000 is a multicenter, open label, dose discovery, phase 1 study to determine safety, tolerability, PK, pharmacodynamics and primary efficacy of BMS-986458 in participants with R/R NHL. This study will be performed in part 2. Dose escalation (part a) will evaluate the escalating oral dose of BMS-986458 to evaluate safety, tolerability, and PK, and guide the determination of the Maximum Tolerated Dose (MTD) and/or OBD of BMS-986458 as a single agent or in combination with rituximab in participants with DLBCL and FL. Dose expansion (part B) will further evaluate the safety, primary efficacy and PK of BMS-986458 administered at or below MTD/OBD in one or more expansion cohorts to determine RP2D at one or more doses as a single agent or in combination with an anti-lymphoma agent.

2. Introduction to the invention. BMS-986458 is under development for the treatment of R/R DLBCL and R/R FL.

There are several agents that show promise for treating hematological malignancies. Subjects with R/R NHL and FL represent heterogeneous populations with poor life expectancy. In vitro, BMS-986458 has demonstrated a variety of pharmacological activities including antiproliferative effects and potent induction of apoptosis in diffuse large B-cell lymphoma (DLBCL) cell lines, including acquired doxorubicin-resistant cell lines. BMS-986458 also exhibits strong immunomodulatory activity in normal lymphocytes (such as follicular helper T cell populations). BMS-986458 has shown potent anti-tumor activity in various DLBCL xenograft models in vivo.

3. Physical/chemical and pharmaceutical properties and formulations

3.1 Physical/chemical properties. BMS-986458-01 was selected for dosage form development and is also known as BMS-986458. Drug substances (drug substances) are mixtures of 2 diastereomers (BMT-648592 and BMT-648599). Two of the three stereogenic centers in each isomer have the (R) configuration. The third stereogenic center has the (R) configuration in BMT-648592 and the (S) configuration in BMT-648599. Physical and chemical properties of BMS-986458-01 drug substance are provided in Table 3.1-1.

TABLE B3-3.1-1 physical and chemical Properties

3.2 Pharmaceutical properties and formulations

3.2.1 Description of dosage forms

20 Mg and 80mg BMS-986458-01 film coated tablets. BMS-986458-01 film coated tablets were used in clinical studies. 20 mg and 80mg film coated tablets are described in Table 3.2.1-1.

20 Tablets of the mg and 80 mg size contained BMS-986458-01 Spray Dried Dispersion (SDD) and inactive ingredients of anhydrous lactose, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, sodium stearyl fumarate and Opadry film coatings. BMS-986458-01 SDD contains 35% w/w of BMS-986458-01 drug substance, which is prepared by spray drying a solution of drug substance and Eudragit L100-55 in methylene chloride and methanol. Tablets are supplied in tightly closed High Density Polyethylene (HDPE) bottles with desiccant and child-resistant closure.

TABLE B3.2.1-1 BMS-986458-01 description of film coated tablets

3.2.2 Pharmaceutical product preparation

20 Mg and 80 mg BMS-986458-01 film coated tablets. For BMS-986458-01 film coated tablets, no preparation is necessary.

3.2.3 Recommended storage and use conditions

20 Mg and 80 mg BMS-986458-01 film coated tablets. BMS-986458-01 film coated tablets should be stored in a tightly closed HDPE container at 2 to 8oC (36 to 46F) protected from light.

4. Non-clinical study. A series of non-clinical primary and secondary pharmacological, safety pharmacological, PK and toxicology studies were performed with BMS-986458. A summary of these studies is provided in the following section.

4.1 Non-clinical pharmacology

4.1.1 Principal pharmacodynamics

4.1.1.1 In vitro pharmacology

Dynamics of endogenous degradation in the OCI-LY-1 model. BMS-986458 is a potent and highly selective CRBN dependent LDD against the oncogenic target BCL 6. BCL6 protein levels were measured using the MSD-ECL assay, and half maximal degradation concentration (DC 50) was determined to be 0.12 nM ±0.0018 after exposure for up to 4 h in different in vitro models (fig. 1; fig. 2A and fig. 2B). DC50 was independently assessed in the individual non-stereoisomers BMT-648592 and BMT-648599. After 2 h incubation, BCL6 degradation characteristics were similar, with BMT-648599 showing a 20-fold difference in potency compared to BMT-648592 (dc50=1.6±1.1 nM compared to 0.08±0.06 nM, respectively) (dose response from representative experiments shown in fig. 2C). The antiproliferative response of both isomers was evaluated and demonstrated that despite the change in degradation potency at 2 h, there was no significant difference in effect on proliferation between the isomer and diastereomer mixture over a period of 5 days, which is likely the result of epimerization to equivalence ratio under experimental conditions (fig. 2D).

In the OCI-LY-1 model, which expressed high levels of BCL6 protein due to THL classification, a single 10 nM dose of BMS-986458 degraded greater than 90% of BCL6 protein in less than 0.5 h and maintained the protein at maximally reduced levels for at least 5 days as assessed by western blot (fig. 2F and 2G).

The antiproliferative effect of BMS-986458 exposure was confirmed to be CRBN dependent, as no loss of proliferation capacity from 5 days of LDD treatment was observed in OCI-LY-1 ^CRBN-/- relative to vehicle, as determined by CellTiter-Glo positive luminescence cell viability assay (CTG) (FIG. 2H).

Proteomic evaluation in OCI-LY-1 model after 4 h and 24 h exposures. BMS-986458 selectivity was assessed in OCI-LY-1 cells using mass spectrometry whole proteomic analysis. BCL6 is the only substrate of significant degradation after 4 h exposure using 100 nM BMS-986458. Above the statistical threshold, no historical new substrate was detected that was affected by CRBN-mediated degradation agents. Evaluation of BMS-986458 under the same conditions at 24 h demonstrated that the abundance of many proteins known to be directly regulated by BCL6 was significantly altered, such as to inhibit the cell cycle checkpoint inhibitor p27/CDKN1B and the regulatory phosphatase SHP-1/PTPN1 ^12,13 (fig. 3).

Antiproliferative and pro-apoptotic responses in BMS-986458 DLBCL cell line models. After exposure to BMS-986458 dose range spanning 0.015-100 nM, the antiproliferative response of twenty DLBCL cell line models was evaluated and evaluated using CTG, after which the response depth was quantified using area under the curve (AUC). This evaluation was initially performed after 120 h incubation with BMS-986458. Under these conditions, 3 out of 20 cell lines (15%) showed more than twice the loss of proliferative capacity (AUC < 5 μm.h) and were classified as acute sensitised. A milder response was observed in the other 15% cell line model (AUC < 7 μm.h). This evaluation was also performed after prolonged LDD exposure covering a 5 day pre-incubation period with 10 nM BMS-986458, previously demonstrated maintenance of 100% BCL6 loss for 120 h (fig. 4). The effect of the initial 5-day reaction was measured and included in the T120/T0 DMSO CTG signal correction factor. After a total of 240 h exposures, the other ten lines exhibited AUC < 7 μm.h and were classified as late response. The total response rate to BMS-986458 within 10 days was 65%, covering both acute sensitive and delayed reactions. The apoptotic response of ten response lines (acute and delayed) was further assessed by annexin v+draq+ staining. After 5 days BMS-986458 treatment, only acute response lines were observed to induce apoptosis (fig. 5), indicating that the delayed response was predominantly cytostatic.

The effect of baseline BCL6 substrate expression on BMS-986458 reaction depth was evaluated. Western blot protein analysis was performed on a large set of cell line models and BCL6 expression was quantified. This was found to be relevant when compared to the BMS-986458 10 day response AUC, corresponding to a R ² pearson correlation coefficient of 0.416. Notably, several cell line models showed significantly higher sensitivity than expected in view of the relatively low BCL6 baseline expression (fig. 6).

In vitro response to BMS-986458 in an engineered doxorubicin resistance model. Apoptosis response to BMS-986458 was evaluated in three paired DLBCL engineered cell line models, which demonstrated acquired doxorubicin resistance, ¹⁴ obtained after incubation with sublethal doses for several months. After 5 days of assay, the proliferation IC50 of doxorubicin was about 1 μm as expected. However, the response to BMS-986458 was significantly altered (fig. 7). In SUDHL-4 and U2932, AUC was reduced by at least 30%, indicating that these models show increased sensitivity to BCL6 degradation. In all doxorubicin resistance (DoxR) models (including WSU-DLCL-2-DoxR, where the proliferative response to BMS-986458 was more resistant than the parent), there was a more than two-fold increase in apoptosis induction, as assessed by AUC. This indicates a significant altered dependence on BCL6 and its pathways of activation after degradation in these models. Western blot protein analysis of the short exposure time course was used to confirm target degradation and evaluate key BCL6 downstream biomarkers (fig. 7). Analysis of baseline MDR1 expression demonstrated that this membrane transporter is the primary driver of doxorubicin resistance, also resulting in impaired BCL6 degradation. Despite this reduced degradation response, several canonical BCL6 downstream biomarkers are still regulated in the DoxR line, including p27/CDKN1B and SHP-1/PTPN6. However, major changes were observed in baseline expression and proliferation signaling activation (including constitutive activity).

AKT ^ser473 (present due to loss of PTEN expression) in SUDHL-4 ^DoxR was then down-regulated after BMS-986458 treatment. The common observation in all DoxR models was that aberrations involved in the anti-apoptotic protein Bcl-2, resulting from either loss of baseline expression or BMS-986458-dependent induction attenuation. Taken together, this in part accounts for the general shift to apoptotic responses following treatment of BMS-986458 in DoxR compared to the parental cell line model.

4.1.1.2 Pharmacological in vivo. The objective of in vivo pharmacological studies was to determine the extent of BCL6 degradation activity and subsequent single agent antitumor activity of BMS-986458 in a cell line derived DLBCL xenograft model. These studies included PK and PD studies in OCI-LY-1 and WSU-DLCL-2 subcutaneous xenograft models to determine the kinetics of intratumoral degradation of BCL6 associated with plasma and tumor BMS-986458 concentrations, as well as single agent antitumor activity.

Pharmacokinetic, pharmacodynamic and anti-tumor activity of BMS-986458 in OCI-LY-1 xenograft models. To evaluate the pharmacodynamic effects of BMS-986458 in vivo, western blot was used to determine and quantify intratumoral levels of BCL6 in an OCI-LY-1 xenograft model in short term 3 or 5 day PK/PD studies with once-a-day (QD) or BID dosing. The PK/PD relationship of BMS-986458 exposure to BCL6 degradation (PD marker) in plasma and tumor was established. The PK/PD relationship was studied at 30 and 60 mg/kg. Dose and time dependent exposure of BMS-986458 and concomitant degradation of BCL6 was observed in OCI-LY-1 tumors. Some variability in PK was observed for the same dose, but PK and PD correlated with each other.

Multiple PK/PD studies were designed to evaluate PK and PD parameters for BMS-986458 administered at 30 mg/kg BID for 5 consecutive days (study AP 9506) ¹⁵ or 3 consecutive days (study AP 9721) ¹⁵ or 60 mg/kg QD for 3 consecutive days (study 06E 26-0023) ¹⁵. Plasma and tumor BMS-986458 concentrations were evaluated along with kinetics of degradation of BCL 6. Significant degradation of BCL6 in tumors (90% to 99%) was observed after five consecutive days of twice daily dosing at 30 mg/kg (fig. 8). During this time, plasma BMS-986458 concentration ranges between 0.00773 and 5.25. Mu.M. Tumor BMS-986458 concentrations ranged between 0.0354 and 0.873. Mu.M (Table 4.1.1.2-1). Similarly, there was significant degradation of intratumoral BCL6 (97% to 98%) when plasma BMS-986458 concentrations ranged between 2.39 and 0.809 μm after three consecutive days of twice daily dosing (fig. 8 and table 4.1.1.2-1). Tumor BMS-986458 concentrations ranged between 1.26 and 0.202. Mu.M (Table 4.1.1.2-1). In the case of QD dosing, a significant and sustained degradation (91% to 98%) of intratumoral BCL6 was observed within 24h, with plasma BMS-986458 concentrations ranging between 11.360 and 0.223 μm. Tumor BMS-986458 concentration range between 3.243 and 0.043. Mu.M. 30 Both mg/kg BID (AP 9721) ¹⁵ and corresponding QD 60 mg/kg (06E 26-0023) ¹⁵ doses resulted in similar levels of BCL6 degradation.

TABLE B3-4.1.1.2-1 plasma/tumor exposure and percent BCL6 degradation of BMS-986458 in OCI-LY-1 xenografts after BID X5, BID X3 or QD X3 administration

Abbreviations NA, inapplicable, BCL6, B cell lymphoma 6;h, hours, BIDx5, twice daily dosing for 5 consecutive days, BIDx3, twice daily dosing for 3 consecutive days, QDx3, once daily for 3 consecutive days, SD, standard deviation.

^a The average BCL6 degradation in BMS-986458 treated tumors was determined as compared to the average vehicle tumor BCL6 levels.

In summary, data from PK/PD studies in OCI-LY-1 tumor models indicate that plasma and tumor BMS-986458 exposure is variable between studies, and this may be due to batch-to-batch variation and compound form. With variations, plasma BMS-986458 concentrations > 2.4. Mu.M at 10 h post-dose will result in continuous tumor BMS-986458 concentrations > 0.04. Mu.M in the case of BID dosing. These exposures resulted in > 90% of BCL6 degradation in OCI-LY-1 tumors. Tumor BCL6 degradation in excess of 97% can be achieved when plasma and tumor concentrations are maintained > 0.8 μm and > 0.2 μm, respectively.

Individual isomers were also tested in pharmacokinetic and pharmacodynamic studies. The individual diastereomers showed equivalent activity ³ in all in vitro pharmacological tests performed. In vivo OCI-LY-1 xenograft models, similar tumor BCL6 degradation levels (OG 00457) ¹⁵ were observed for BMS-986458 and its individual diastereomers at the same dose level. Additional chiral analysis of the exposure results also showed significant epimerization in mice, with BMT-648599 being slightly more prone to epimerization in mice than BMT-648592.

BMS-986458 was tested for single agent antitumor activity (AP 9553) ¹⁵ in the OCI-LY-1 xenograft model with 15, 30 and 60 mg/kg BID dosing. In this model, a single cycle CHOP therapy (combination of cyclophosphamide-day 1 single dose, doxorubicin-day 1 single dose, vincristine-day 1 single dose, and prednisone-day 1 to day 5 QD dosing for 5 doses [ QD x 5 ]) was used as a positive control. BMS-986458 was observed to have significant (p < 0.0001) dose-dependent antitumor activity with tumor volume reductions of 53%, 62% and 76% at 15, 30 and 60 mg/kg BID, respectively. In the case of BMS-986458 at all doses tested in this response study, at time points 2h and 10 h after the last dose, plasma and tumor samples collected at the end of the response study showed deep (95% to 99%) and statistically significant (p < 0.0001) BCL6 degradation. Pharmacokinetic data from these samples indicate that sustained degradation of BCL6 ≡97% (which can be achieved at > 0.24 μm plasma and > 0.15 μm tumor BMS-986458 concentration after 10 h doses twice a day) confers efficacy in excess of 62% TVR in the OCI-LY-1 xenograft model.

Table B3-4.1.1.2-2 percent tumor volume reduction in OCI-LY-1 xenografts, plasma/tumor exposure to BMS-986458, and percent BCL6 degradation (AP 9553) after twice daily dosing for 19 consecutive days

In another study (AP 9742) ¹⁵, BMS-986458 and its isoforms BMT-648592 and BMT-648599 were evaluated for anti-tumor activity in an OCI-LY-1 xenograft model at 30 and 60 mg/kg BID dosing. BMS-986458 and both isoforms showed significant antitumor activity when compared to vehicle controls, with TVR of BMS-986458 at 60% and 64%, TVR of BMT-648592 at 30 and 60 mg/kg, respectively, 48% and 83%, and TVR of BMT-648599 at 54% and 66% (OG 00457) ¹⁵. No significant difference in antitumor activity of BMT-648599 was observed at a dose of 30 mg/kg compared to BMS-986458. The antitumor activity of BMT-648592 at higher doses (60 mg/kg) was significantly (p < 0.01) better than BMS-986458 at the corresponding dose level. The extent of BCL6 degradation was also assessed from samples collected from the group dosed with BMS-986458 and isomers (BMT-964592 and BMT-964599). Similar to BMT-986458, deep degradation of BCL6 was observed in OCI-LY-1 tumors with the isomers BMT-648592 and BMT-648599. Similar degradation depths (94% to 100%) were observed at time points 6, 10 and 14 h for all three compounds. Additional chiral analysis of the exposure results also indicated that significant epimerization occurred in mice. Overall, plasma and tumor BMS-986458 concentrations and anti-tumor activity in this OCI-LY-1 study appear to be moderately different compared to other OCI-LY-1 xenograft studies. The anti-tumor response with the test compound or positive control (CHOP therapy) was lower in the current study than in the other study (OG 00457) ¹⁵ at the corresponding dose level.

BMS-986458 was tested in the OCI-LY-1 xenograft model at 15, 45 and 60 mg/kg BID in a subsequent dose response study (AP 9776) ¹⁵. Significant (p < 0.0001) single agent dose-dependent antitumor activity was observed at 15 mg/kg and 45 mg/kg, with a 67% and 84% reduction in tumor volume, respectively. Similar antitumor activity (84% TVR) was observed with BMS-986458 dosed with 45 and 60 mg/kg BID (FIG. 9 and Table 4.1.1.2-3). The minimum effective dose in this study (as defined by a reduction in tumor volume of about 65%) was a BID dose level of 15 mg/kg.

Consistent with findings from AP9553 and AP9742 ¹⁵, deep and statistically significant BCL6 degradation was observed in OCI-LY-1 tumors collected at the end of the study. At time point 6 h, 98%, 99% and 99% BCL6 degradation were achieved with BMS-986458 at 15, 45 and 60 mg/kg BID dose, respectively. In addition, 98%, 100% and 99% BCL6 degradation was observed at time points 10 h with BID doses of 15, 45 and 60 mg/kg, respectively (fig. 10). At time points 14 h, 95%, 100% and 100% BCL6 degradation was observed with BMS-986458 at 15, 45 and 60 mg/kg BID doses, respectively (table 4.1.1.2-3). This pharmacodynamic data, along with plasma and tumor BMS-986458 concentration analysis, showed that BMS-986458 at all three dose levels used in this study resulted in near complete BCL6 degradation (95% to 100%). The calculated plasma AUC _(0-24 _h) values for each BMS-986458 dose group were 26.1, 137.2 and 256. Mu.M ∙ h/mL for 15, 45 and 60 mg/kg BID, respectively (Table 4.1.1.2-3). Continuous degradation of > 95% is necessary to confer > 65% response at plasma and tumor continuous concentrations > 0.07 and > 0.04 μm BMS-986458, respectively. BMS-986458 was well tolerated during treatment and there was no significant weight loss at the dose levels tested.

In this study, the antitumor activity of BMS-986458 was compared to BMT-648592, the individual isoforms administered with 15, 45 and 60 mg/kg BID. In the case of BMT-648592 at 15, 45 and 60 mg/kg, the tumor volume reduction was 66%, 81% and 81%, respectively. No significant difference in antitumor activity was observed between BMS-986458 and the individual isomer BMT-648592 (Table 4.1.1.2-3). Similar to BMT-986458, deep degradation of BCL6 was observed with individual isomer BMT-648592 in OCI-LY-1 tumors collected at the termination of the study. Similar degradation depths (94% to 100%) were observed at time points 6, 10 and 14 h for both compounds (table 4.1.1.2-3; og00457 ¹⁵).

In summary, the data from the study using the OCI-LY-1 xenograft model combined with PK and PD data obtained from samples collected at the end of the study, suggest that plasma and tumor BMS-986458 exposure and anti-tumor activity at a given dose are variable between studies, and this may be due to batch-to-batch variation and form of the compound. Considering the variation, a sustained degradation of tumor BCL6 degradation > 95% appears to be necessary to show a response of about 65% TVR (except for study AP9742 where TVR was lower in all study groups including positive control). This degree of tumor BCL6 degradation and corresponding anti-tumor response can be achieved at 10: 10 h post-dose with repeated BID dosing of 15-30 mg/kg at plasma and tumor BMS-986458 concentrations > 0.24 and > 0.15 μm, respectively. With 15 mg/kg BID, the calculated BMS-986458 effective plasma AUC (0-24) h from study AP9776 was 26.1 μ M.h/mL (Table 4.1.1.2-3).

Table B3-4.1.1.2-3 plasma exposure and percent BCL6 degradation in OCI-LY-1 xenografts after twice daily dosing for 21 consecutive days (AP 9776)

Anti-tumor Activity of BMS-986458 in WSU-DLCL-2 xenograft model (AP 9664) with twice daily dosing. BMS-986458 was tested for antitumor activity of a single agent in a WSU-DLCL-2 xenograft model with 30 and 60 mg/kg BID dosing. In this model, a single cycle CHOP therapy (combination of cyclophosphamide-day 1 single dose, doxorubicin-day 1 single dose, vincristine-day 1 single dose, and prednisone-day 1 to day 5 QD dosing for 5 doses [ QD x 5 ]) was used as a positive control. BMS-986458 significantly inhibited the growth of WSU-DLCL-2 xenograft tumors, with 69% and 74% TVR at 30 and 60 mg/kg BID, respectively (Table 4.1.1.2-4). No significant (p > 0.05) weight loss was observed in the 30 mg/kg BMS-986458 treated group. On day 29 (p < 0.01), 33 (p < 0.05), 36 (p < 0.05) and 39 (p < 0.01), small but statistically significant weight loss was observed in animals treated with 60 mg/kg BMS-986458.

Similar to the data in the samples collected at the termination of the study from the study in the OCI-LY-1 xenograft model (study AP9553, AP9742 and AP 9776), deep and statistically significant BCL6 degradation was observed in WSU-DLCL-2 tumor lysates from the group treated with BMS-986458 compared to the vehicle group. At 30 mg/kg, there was 98%, 97% and 86% BCL6 degradation at 6, 10 and 14 h. At 60 mg/kg, at 6, 10 and 14 h, BCL6 degraded to 99%, 99% and 98% (table 4.1.1.2-4). This pharmacodynamic data, as well as plasma and tumor BMS-986458 concentration analysis, showed that 30 and 60 mg/kg BID administration resulted in almost complete BCL6 degradation (97% to 99% except 14 h at 30 mg/kg, which showed 86% degradation). Continuous plasma and tumor BMS-986458 concentrations > 0.1 and > 0.07. Mu.M, respectively, are necessary to confer > 65% anti-tumor response.

Table B3-4.1.1.2-4 plasma/tumor exposure and percent BCL6 degradation (AP 9664) of BMS-986458 in WSU-DLCL-2 xenografts after twice daily dosing for 14 consecutive days

In summary, data from studies using the WSU-DLCL-2 xenograft model combined with PK and PD data obtained from samples collected at the end of the study showed that > 95% tumor BCL6 degradation could be achieved with plasma and tumor BMS-986458 concentrations > 0.7 and > 0.5 μm, respectively, and a minimum effective dose to > 65% TVR of 30 mg/kg, BID.

4.1.2 Secondary pharmacodynamics. The effect of BMS-986458 treatment on BCL6 degradation was evaluated in normal immune populations associated with BCL6 functional dependence or commonly observed sensitivity to treatment.

4.1.2.1 Effect of BMS-986458 on follicular helper T cells. The effect of BMS-986458 was evaluated in T-follicular helper cells in vitro. These populations are not normally present outside the germinal center, and therefore, evaluation requires differentiation from naive T cells isolated from healthy donor PBMCs. Under the conditions of these studies, maximum induction of cxcr5+pd1+icos+btla+ from sorted cd4+ cells was observed 4 days after stimulation with anti-CD 3/CD28 beads, activin a and IL-12, with differentiation rates consistently between 20% and 30%. Further characterization of Tfh cells differentiated on day 4 showed strong induction of canonical Tfh cytokine IL-21 and induced expression of BCL6 (fig. 11C). When 10 nM BMS-986458 is included with stimulation, the 4 th day differentiation rate of cxcr5+pd1+icos+btla+il-21 secreting cells relative to vehicle is unaffected despite complete degradation of BCL6 and this is consistent between donors. However, IL-10 secretion was significantly increased (FIG. 11F). When analyzing cell phenotypes on days 8 and 12, there was a BMS-985458 dose-dependent loss of cxcr5+pd1+ iTfh cells between donors and was accompanied by a loss of IL-21 secretion without significant loss of viability. Phenotypic identification of these cells was performed after BMS-986458 treatment.

4.1.2.2 Effect of BMS-986458 on neutrophil maturation. Cell viability and proliferation of cd34+ derived cells was unaffected by BMS-986458 exposure. At doses exceeding clinically relevant concentrations, a slight decrease in neutrophils was observed, reflecting maturation stage II. This slight effect is reversible, as complete recovery is observed after compound elution.

4.2 Non-clinical pharmacokinetics and metabolism. In vitro and in vivo studies were performed to characterize the absorption, PK, distribution, metabolism and drug interaction potential of BMS-986458. Robust and reproducible bioassay methods for determining BMS-986458 concentrations were developed and used for PK and Toxicokinetic (TK) studies. The pharmacokinetics and oral bioavailability of BMS-986458 were evaluated in mice (IV PK only), rats, dogs and monkeys. In vitro studies were performed to assess BMS-986458 metabolism, permeability, plasma protein binding, blood-plasma distribution, inhibition and induction potential of cytochrome P450 (CYP) enzymes, exointestinal transporter substrates and inhibition potential, hepatic and renal transporter inhibition potential, and uridine diphosphate glucuronyl transferase 1A1 (UGT 1 A1) inhibition potential. In addition, circulating metabolites from plasma samples from multiple dose exploratory toxicity studies in rats and dogs were evaluated. Table 4.2-1 lists analytical methods validated, absorbed, distributed, metabolized, PK and drug interaction studies performed to date with BMS-986458.

TABLE B3-4.2-1 non-clinical pharmacokinetic Studies

4.2.1 Biological analysis method. Robust and reproducible bioassay methods for determining BMS-986458 plasma concentrations were developed and used for PK and TK studies. In non-clinical GLP-compliant toxicology studies, liquid chromatography tandem mass spectrometry (LC-MS/MS) bioanalytical assays for quantification of BMT-648592 and BMT 648599 (diastereomers of BMS-986458) were fully validated ^33,34 according to the health agency guidelines (Health Authority Guidance). Details of the validated methods are summarized in Table 4.2.1-1. Other PK and drug metabolism studies use bioanalytical methods suitable for the purpose to quantify BMS-986458.

Table B3-4.2.1-1 biological analysis method for plasma concentration measurement of BMS-986458

4.2.2 Absorption and pharmacokinetics. BMS-986458 was evaluated for PK in male CD-1 mice (IV only), male Sprague Dawley rats, male beagle dogs and male cynomolgus monkeys after Intravenous (IV) infusion and oral administration. The average PK parameters for IV and oral administration are summarized in table 4.2.2-1.

Following a single IV infusion, BMS-986458 systemic plasma clearance (CLTp) was low (+.11% of liver blood flow) in mice and dogs, and moderate (+.44% of liver blood flow) ³⁵, 36 in rats and monkeys. The steady state distribution volume (Vss) ranged from 0.217 to 2.45L/kg, and was low in mice and dogs (0.4 to 0.7 times the total body water), low to medium in rats (0.4 to 3.0 times the total body water) and medium in monkeys (2.4-3.5 times the total body water), indicating that BMS-986458 had some extravascular distribution. The apparent terminal HALF-life (T-HALF) of BMS-986458 was medium and ranged from 0.9 to 5.5 hours in all non-clinical species tested. ²⁰ ,21,22,23

BMS-986458 was absorbed after a single oral dose, with average Tmax values ranging from 2.67 to 4.0 hours in rats, dogs and monkeys. BMS-986458 had moderate oral bioavailability (22% to 37%) among all 3 non-clinical species tested. ²⁰ ,21,22,23

Furthermore, after a single IV infusion or oral dose of the individual diastereomers of BMS-986458 (BMT-648592 or BMT-648599), significant interconversions between the diastereomers (up to 41% based on AUC) were observed in rats, dogs and monkeys. ²⁰

The toxicology ⁴⁵ ^, ⁴⁷ of BMS-986458 was examined after multiple oral doses in toxicology studies. In general, a large inter-individual variability in exposure was observed in both rats and dogs. After repeated administration of BMS-986458, the average BMS-986458 systemic exposure (AUC [0-24h ]) in rats was generally less than dose-proportional increase in the dose range of 50 to 500 mg/kg BID, given the large inter-individual variability, and without significant sex differences. In rats, the average AUC (0-24 h) values at day 28 were slightly lower (0.4 to 0.8×) at day 1. In dogs, the average AUC (0-24 h) values after repeated dosing were generally greater than dose-proportional increases from 5 to 15 mg/kg BID and approximately dose-proportional increases from 15 to 50 mg/kg BID, and given the large inter-individual variability, no significant sex differences and no accumulation or loss of exposure were noted.

Overall, BMS-98458 exhibited low to moderate systemic clearance, low to moderate steady state distribution volumes, and acceptable oral bioavailability in mice (no bioavailability data available), rats, dogs, and monkeys in preclinical species. Significant interconversion between diastereomers (up to 41% based on AUC) was observed following IV infusion or PO administration of individual diastereomers in rats, dogs and monkeys.

4.2.3 Distribution. BMS-986458 was evaluated for in vitro plasma protein binding in CD-1 mice, sprague-Dawley rats, new Zealand white rabbits (female), beagle dogs, cynomolgus monkeys, and human (pooled male and female) plasma. Plasma protein binding at 0.1, 1 and 10 μg/mL was assessed in plasma of each species using an ultracentrifugation method. Furthermore, the in vitro blood-plasma distribution of BMS-986458 was evaluated at 1 μg/mL in mouse, rat, rabbit, dog, monkey and human whole blood.

In mouse, rat, rabbit, dog, monkey and human plasma, BMS-986458 binds to plasma proteins up to 99.95%, 99.84%, 99.78%, 99.99%, 99.68% and 99.06%, respectively, with corresponding unbound fractions (fu) < 1%. This result is consistent with earlier exploratory results obtained from modified rapid equilibrium dialysis (10-fold plasma dilution) in combination with exploratory bioanalytical methods. No significant difference in plasma protein binding was observed over the concentration range tested in all species, indicating that binding to plasma protein was not saturated in all species. Taking into account the uncertainty of plasma protein binding measurements (i.e., non-measurable concentrations in multiple protein-free supernatant samples), orthogonal assays (i.e., flux dialysis methods) were performed and plasma fu < 1% of BMS-986458 was confirmed in all species tested. In addition, a species difference of fu (5.3×) was observed between humans and dogs, whereas no substantial difference (+.2×) was observed between humans compared to rats or rabbits. ²⁰ ,25

In mice, rats, rabbits, dogs, monkeys, and humans, the blood-plasma concentration ratios of BMS-986458 were 0.68, 0.87, 0.62, 0.74, and 0.76, respectively, indicating limited partitioning of BMS-986458 into blood cells. ²⁵

The distribution of BMS-986458 in the brain was determined in rats after a single oral dose ranging from 10 to 300 mg/kg. After a single oral dose of 10 to 300 mg/kg to Sprague-Dawley rats, brain BMS-986458 concentration was ∈0.22 μΜ or less in female rats 24 hours post-dose (brain/plasma ratio ∈0.10 at ∈100 mg/kg and not properly assessed at ∈30 mg/kg due to low or non-measurable [ i.e., < LLOQ 0.0029 μΜ ] concentration) and not measurable in male rats 48 hours post-dose, indicating minimal penetration across the blood brain barrier. In addition, BMS-986458 is also distributed in the liver, spleen, lymph nodes, ileum, heart and lung, with tissue/plasma ratios ranging from 0.6 to 42 (see section 4.3.1.1 for details non-clinical toxicology). ²⁶

The distribution of BMS-986458 in other tissues and the transport of BMS-986458 across the placental barrier have not been evaluated.

4.2.4 Metabolism

4.2.4.1 Metabolic stability. The metabolic stability ²⁰ ^, ²⁴ of BMS-986458 was evaluated with NADPH-fortified liver microsomes and hepatocytes from humans, rats, mice (microsomes only), dogs and monkeys. BMS-986458 (0.5. Mu.M) was incubated with NADPH-fortified liver microsomes (1 mg/mL) at 37℃for up to 45 minutes to determine the in vitro half-life and rate of CYP-mediated metabolism. After the incubation period, the mixture was precipitated with acetonitrile and centrifuged. The supernatant was analyzed by LC-MS/MS to obtain the relative amount of BMS-986458 remaining in the reaction mixture. In addition, BMS-986458 (1.0. Mu.M) was incubated with a pooled cryopreserved hepatocyte suspension (1X 10 ⁶ cells/mL) of rats, dogs, monkeys and humans for up to 120 minutes at 37℃to determine in vitro half-life. The reaction was stopped by adding acetonitrile containing an internal standard (i.e., terfenadine) and centrifuged. The supernatant was analyzed by LC-UV [ ultraviolet ]/MS and the residual amount of BMS-986458 in the reaction mixture was quantified against a standard curve.

A variable rate of CYP-mediated metabolism (0.0070 to 0.0189 nmol/min.mg protein) of BMS-986458 was observed with liver microsome incubation. The highest metabolic rate was observed in monkeys and rats, followed by humans and mice, and the lowest metabolic rate was observed in dogs. BMS-986458 has an in vitro half-life in hepatocytes ranging from 53 to 114 minutes, the longest half-life observed in human hepatocytes.

4.2.4.2 And (5) metabolite spectrum analysis. Following oral administration, metabolism of BMS-986458 was assessed in mice, rats, dogs, monkeys and human hepatocytes, as well as in plasma samples from rats and dogs. BMS-986458 (2. Mu.M) was incubated with cryopreserved hepatocytes from animals (pooled males) and humans (males alone) for up to 4 hours at 37 ℃. After the incubation period, the mixture was precipitated with acetonitrile and centrifuged. Supernatants were analyzed by LC-MS/MS with on-line UV for metabolite identification.

In human hepatocytes, BMS-986458 is metabolized primarily by demethylation and hydrolysis and oxidation, followed by less-active glucuronidation. Additional secondary metabolic pathways in human hepatocytes include monooxidation, dehydrogenation, and cleavage of the linker between the piperidine and indazole moieties. The most prominent metabolites in human hepatocytes (detectable by UV) are M11 (demethylation), M6 and M8 (hydrolysis), M9, M10, M13a and M13b (monooxidation), M21 (oxidation followed by glucuronidation), M20 (cleavage of the linker between piperidine and indazole moieties), M12a and M12b (dehydrogenation). Of these metabolites, all metabolites were detected by UV in hepatocytes of mice, rats, dogs and monkeys, except that M9, M13 and M13b were detected by MS only in hepatocytes of dogs, as was M13b in hepatocytes of monkeys. Although there were quantitative differences in metabolite formation between species, qualitatively, all metabolites formed in human hepatocytes were also formed in hepatocytes from rats and/or dogs (both species for preclinical safety testing), indicating the absence of unique human metabolites. ³⁷

In plasma from rats and dogs after multiple oral administrations, BMS-986458 was the predominant (76% to 90%) drug-related circulating species, and the observed metabolites were similar to those observed in hepatocytes, indicating good in vitro and in vivo correlations. The metabolites detected by UV in rat plasma were M9, M9a, M10, M11, M13a and M20, and in dog plasma M6, M9a, M10, M11, M12a, M12b, M13a, M13b, M16 and M20 were detectable by UV. ²⁸ ^, ²⁹

4.2.4.2 Reactive phenotyping. Phenotypic studies of human CYP isoforms and human liver microsomes with isozyme specific inhibitors indicate that BMS-986458 is metabolized primarily by CYP3A4/5 and that CYP1A2, CYP2C8, CYP2C9 and CYP2C19 are less involved. ²⁷

4.2.5 And (5) excretion. No study has been performed to investigate the excretion of BMS-986458. The excretion of BMS-986458 or its related components into breast milk has not been evaluated.

4.2.6 Pharmacokinetic drug interactions

4.2.6.1 Evaluation of drug interactions based on metabolism. In vitro studies were performed to assess the likelihood of drug-drug interactions (DDI) due to BMS-986458 inhibiting CYP isozymes. Under experimental conditions used to examine direct inhibition, BMS-986458 had no inhibitory effect on CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 (IC 50 > 20 μm). In vitro, BMS-986458 exhibited a time-dependent inhibition of CYP3A4 when using midazolam or testosterone as substrate, respectively, with an IC50 shift > 16.3-fold or > 18.1-fold (not pre-incubated with BMS-986458 compared to BMS-986458). Using midazolam as a substrate, the enzyme inactivation rate (kinact) and inhibition constant (KI) were determined to be 0.054/min and 28. Mu.M, respectively. In addition, a weak time-dependent inhibition of CYP1A2 by BMS-986458 was also observed, IC50 shift > 2.5-fold. Based on expected clinical doses and exposures and interpreting in vitro CYP inhibition data ³⁸, 39 according to FDA and EMA guidelines, there may be a potential clinically relevant systemic DDI if BMS-986458 is co-administered with a strong CYP3A4 inhibitor or inducer, a sensitive CYP3A substrate, or a CYP3A substrate with a narrow therapeutic window. The potential of DDI will be evaluated throughout the development of BMS-986458. In addition, BMS-986458 showed inhibition of Uridine Diphosphate (UDP) -glucuronyl transferase (UGT) 1A1 with an IC50 of 0.15. Mu.M. ²⁴

An in vitro study was performed to determine the potential of BMS-986458 to induce expression of CYP1A2, CYP2B6 and CYP3A4 mRNA in cryopreserved human hepatocyte cultures. Treatment of cultured human hepatocytes with BMS-986458 (0.1 to 20. Mu.M) resulted in a≤4-fold increase in CYP1A2 mRNA relative to vehicle control. mRNA increase was 9.86% of the positive control inducer. These results indicate that BMS-986458 may be a weak CYP1A2 inducer. In contrast, BMS-986458 (0.1 to 20. Mu.M) treatment resulted in < 2-fold increase in CYP2B6 and CYP3A4 mRNA relative to vehicle control, and an increase in mRNA of 4.94% of the positive control inducer, indicating that BMS-986458 is not an inducer of CYP3A4 or CYP2B 6. ³⁰

4.2.6.2 Evaluation of drug interactions based on transporters. To assess passive permeability, BMS-986458 was tested in Madin-Darby canine kidney cell line II (MDCK II) in which the endogenous canine multidrug resistance 1 gene (cMDR 1) was knocked out. At a concentration of 3. Mu.M in assay buffer containing 0.5% bovine serum albumin, BMS-986458A to B permeabilities were about 151 nm/sec. However, the a to B permeability is likely to be underestimated due to the noted poor recovery (64%). The passive permeability values observed in the MDCKII-mdr 1-knockout cell assay are consistent with the moderate oral bioavailability of BMS-986458. ²⁴

3. Mu.M BMS-986458 was tested in Caco-2 (with or without efflux transporter inhibitors) and MDCKII cells expressing MDR1 to assess the potential of the efflux transporter substrates. In Caco-2 cells, BMS-986458 recovery was 60% and efflux ratio > 4.4. In addition, 3. Mu.M BMS-986458 was incubated with Breast Cancer Resistance Protein (BCRP) inhibitor (1. Mu.M KO 143) or P-glycoprotein (P-gp) inhibitor (5. Mu.M zoolaquin) in Caco-2 cells. The efflux ratio is only slightly reduced with the addition of BCRP and P-gp inhibitors and remains > 3.1 or > 2.5, respectively. The recovery of BMS-986458 from this assay was relatively low (54% to 63%), indicating that permeability was likely to be underestimated. In MDCK cells expressing MDR1, the efflux ratio was 111, and BMS-986458 recovery ranged from 58% to 110% ²⁴. In summary, in vitro assays showed that BMS-986458 was a substrate for P-gp and most likely BCRP.

In vitro assays were performed in Caco-2 to assess the potential of BMS-986458 to inhibit the efflux transporter. BMS-986458 was identified as an inhibitor of P-gp and BCRP with half maximal inhibitory concentration (IC 50) values of 3.3 and 1.6. Mu.M, respectively. These data indicate that BMS-986458 has the potential to cause inhibition of P-gp and/or BCRP in the gut and that absorption of BMS-986458 may be increased in the presence of P-gp inhibitors. ²⁴

The potential of BMS-986458 (up to 50 μm) to inhibit a set of liver and kidney transporters was evaluated in the everting plasma membrane (vesicle) of HEK293 cells expressing uptake transporters and baculovirus-infected insect cells from expression of human Bile Salt Export Pumps (BSEP) or human multi-drug resistance related proteins (MRP) 2. BMS-986458 shows inhibition of a variety of transport proteins, organic Anion Transport Polypeptide (OATP) 1B1, OATP1B3, MRP2, organic cation transport protein (OCT) 1, OCT2, BSEP, multidrug and toxin efflux proteins (MATE) 1 and MATE2K, wherein the IC50 value ranges from 0.29 to 9.5. Mu.M. Overall, the potential for systemic interactions with drug transporters appears to be low based on the expected initial clinical exposure (within the expected effective dose range). However, since clinical exposure is available, the potential for DDI will be continuously assessed.

4.2.7 Other pharmacokinetic studies. Based on PK profile of non-clinical species, and using the abnormal growth scale corrected for unbound total clearance in mice, rats, dogs and monkeys, low human clearance (3.6 mL/min/kg) was predicted. Human Vss was predicted using the sie-Tozer model ⁴⁰ with steady state distribution volumes (Vss) and plasma protein binding data from rats, dogs and monkeys. BMS-986458 was predicted to have a moderate Vss of 2.08L/kg in humans. The predicted human T-HALF was about 10.6 hours. Because BMS-986458 is highly bound in all species tested (unbound fraction in plasma [ fu, p ] < 1%), the accuracy of human CL and Vss estimates may be affected by protein binding measurements. The predicted human absorption rate constant (ka) and oral bioavailability (F) were 0.32 h ^-1 and 26%, respectively, both of which are geometric averages derived from rats, dogs and monkeys. ³²

These PK parameters resulted in an expected steady state Cmax of 85 ng/mL and AUC (0-12 h) of 754 ng.h/mL at the expected clinical starting dose of 40 mg BID and assuming 70 kg humans in multiple incremental dose studies. At the highest proposed dose of 640 mg BID, these PK parameters resulted in a Cmax of 1354 ng/mL and an AUC (0-12 h) of 12,064 ng.h/mL at steady state. ³²

4.2 Non-clinical toxicology. A series of non-clinical studies have been completed including single dose toxicity, repeat dose toxicity, genotoxicity, safety pharmacology, and phototoxicity to assess the potential toxicological profile of BMS-986458. Rats and dogs were chosen for toxicity studies because BMS-986458 has pharmacological activity in both species. The toxicology evaluation scope of BMS-986458 as outlined in table 4.3-1 supports its proposed clinical testing in phase 1 studies in humans. Key research was conducted in member countries of the institute of economic and development (OECD) data mutual approval program (Organization for Economic Cooperation and Development (OECD) Mutual Acceptance of Data program) and according to OECD test guidelines and GLP guidelines or GLP regulations of the federal regulation (CFR) 21 part 58 (21 CFR 58) of the united states food and drug administration (United States Food and Drug Administration, FDA).

TABLE B3-4.3-1 toxicology studies

4.3.1 Toxicity at Single dose

4.3.1.1 Single dose study in rats. In single dose toxicokinetics and tolerability studies in rats ²⁶, BMS-986458 was administered to a group of 6 rats (3M/3F) by oral gavage at a single dose of 0 (vehicle), 10, 30, 100 or 300 mg/kg. All doses were administered at 10 mL/kg in vehicle/vehicle consisting of 10% Captisol in 25 mM citrate buffer (ph=3). Evaluation criteria included pharmacokinetic, biodistribution, BCL6 expression, survival, body weight, clinical observations, clinical pathology, and cytokine evaluation. These evaluations were performed at designated times during the study dosing. The summary of the toxicology of BMS-986458 is presented in the following table.

TABLE B3-4.3.1.1-1 Poison kinetics summary-BMS-986458 mean values in male and female rats

At 24 or 48 hours post-dose, tissues were collected for bioassays to measure the amounts of BMS-986458 in plasma and tissues and to determine the corresponding tissue/plasma ratios. Plasma and tissue were collected from female rats 24 hours after dosing and from male rats 48 hours after dosing for bioassay. In female rats, the highest drug concentration and tissue/plasma ratio were found in the ileum at all doses. In male rats, the highest drug concentration and tissue/plasma ratio were found in the liver after a dose of 10 mg/kg and in the ileum after a dose of ≡30 mg/kg. Tissue bioanalytical data are summarized in the table below.

TABLE B3-4.3.1.1-2 BMS-986458 concentration in plasma and tissues of female rats 24 hours after administration

Spleen, liver and heart samples were collected from females 24 hours after dosing and from males 48 hours, and BCL6 expression was assessed using western blot and normalized for β -actin expression. Rats showed reduced BCL6 protein in spleen and lung 24 hours or 48 hours after treatment. There was no significant decrease in BCL6 protein in the heart.

BMS-986458 was well tolerated at all doses and had no adverse effects on body weight or clinical signs, no significant changes in serum chemistry or hematology, no findings of macroscopic examination at necropsy. Serum cytokine analysis showed BMS-986458-associated decreases in CX3CL1, IL-12 (p 70) and IL-5, which CX3CL1, IL-12 (p 70) and IL-5 are involved in T-cell and B-cell differentiation and/or cell adhesion/migration activity and may be pharmacologically associated with BCL 6. Additional compound-related changes in IL-12 (p 70), CXCL10, leptin and VEGF-A may be indicative of steady state changes in angiogenic activity.

In summary, BMS-986458 administered in rats at single doses up to 300 mg/kg was well tolerated. Evidence of statistically significant decreases in BCL6 protein levels were detected in the spleen and lung 24 hours and 48 hours post-dose, indicating pharmacological activity in these tissues.

4.3.1.2 Maximum tolerated dose and toxicodendron assessment of dogs administered a single oral dose. In the single dose maximum tolerated dose study ⁴¹, BMS-986458 was administered to groups of 2 dogs (1 male and 1 female/group) by oral gavage at doses of 0 (vehicle), 15, 45 or 90 mg/kg. Evaluation criteria included pharmacokinetic, proteomic, survival, body weight, clinical observations, food consumption, and clinical pathology assessment. Food consumption was assessed on days 1 to 3. Clinical pathology was assessed prior to testing and on days 3 and 6. Pharmacokinetic was assessed at 0.5, 1, 4, 5, 7, 24 and 48 hours post-dose.

No dose proportionality was observed between the 15 mg/kg to 90 mg/kg doses. Plasma concentrations of BMS-986458 individual diastereomers (BMT-648592 and BMT-648599) have a significant trend toward BMT-648592 at a ratio of 2:1. There was a large individual variability between 2 dogs dosed with 45 mg/kg or 90 mg/kg. Individual toxicological data for BMS-986458, BMT-648592 and BMT-648599 are summarized below.

TABLE B3-4.3.1.2-1 summary of toxicology-evaluation of maximum tolerated dose and toxicology for dogs administered a single oral dose

Proteomic evaluation of leukocytes collected from peripheral blood showed 85% to 90% BCL6 degradation in all dose groups at 4 hours post-dose. At all doses, there was a partial rebound of BCL6 levels 24 hours post-dose, with 70% to 80% degradation lasting at 90 mg/kg. BCL6 was selectively degraded by BMS-986458 at 4 hours post-dose compared to vehicle in all dose groups. In this comparison, no additional proteins exhibited statistically significant modulation, including canonical CELMoD substrates (e.g., IKZF1, IKZF3, GSPT1, ck1α). No other proteins were significantly regulated 24 hours after dosing, indicating that the duration of BCL6 degradation may be insufficient to see its secondary effects of degradation in leukocytes.

Dogs in all dose groups appeared normal (alert and active) when observed before and up to 6 days after BMS-986458 administration. There were no clinical signs, effects on body weight or food consumption. An increase in WBCs and neutrophils was observed at all doses compared to before the test. In female dogs at 90 mg/kg, elevated alanine Aminotransferase (ALT) levels were noted as compared to before testing and continued until day 10 post dosing (up to 2.02 times on day 10).

In summary, BMS-986458 administered at a single oral dose of 15, 45 or 90 mg/kg, respectively, did not reach dose-proportional exposure, with a plasma concentration ratio of 2 diastereomers BMT-648592 and BMT-648599 of 2:1. At 4 hours post-administration BCL6 degradation was 85% to 90%, whereas by 24 hours there was dose-related BCL6 rebound, with only 90 mg/kg dose maintaining at least 70% BCL6 degradation. BMS-986458 dose administration showed good selectivity in terms of BCL6 degradation and no noticeable off-target activity. BMS-986458 is well-tolerated, wherein only high-dose females (4201) exhibit elevated ALT levels for at least 10 days after administration (AUC [ INF ] = 1512 μg.h/mL).

4.3.2 Repeat dose toxicity

4.3.2.1 5 Day non-GLP study in rats. In the exploratory 5-day toxicity study ⁴², BMS-986458 was administered to a group of 6 male rats (except on day 5, where rats received only a single dose in the morning) by oral gavage BID (8±0.5 hours apart) at a dose of 0 (control), 50, 150, or 500 mg/kg (total daily dose of 0, 100, 300, and 1000 mg/kg/day). All doses were administered at 10 mL/kg BID in vehicle consisting of 0.25% Methocel E4M in water. Endpoints include survival, toxicology, metabolite profiling, clinical observations, body weight, food consumption, clinical pathology (hematology, coagulation, clinical chemistry and urinalysis) evaluation, pharmacodynamics (BCL 6 expression) evaluation, organ weight, and gross and microscopic pathology analysis. Scheduled necropsies (6/group) were performed at the end of the 5 day dosing period.

On day 4, the average systemic exposure (AUC [0-24h ]) of BMS-986458 and its diastereomers BMT-648592 and BMT-648599 generally increased in a dose-proportional manner from 50 to 150 mg/kg BID and less than in a dose-proportional manner from 150 to 500 mg/kg BID after repeated dosing. A summary of the toxicology of BMT-986458 and diastereomers BMT-648592 and BMT-648599 is presented in the following table.

Table B3-4.3.2.1-1 summary of toxicology-average-five day oral exploratory toxicity study in Male rats

^a BMS-986458 is a mixture of BMT-648592 and BMT-648599. The pharmacokinetic parameters were estimated from plasma BMS-986458 concentration profiles, which were constructed by summing the individual BMT-648592 and BMT-648599 concentrations at each time point.

Source research DN22115 ⁴²

The metabolite profile of the rat plasma samples showed no change in BMS-986458 as the primary drug species in circulation, which represents about 76% of the total circulating species in rat plasma, after multiple daily doses of 500 mg/kg BID. The most abundant metabolites in rat plasma are formed via the monooxidation of the parent (M9 and M10), accounting for 10.9% and 6.9% of the total circulating species. Low levels of oxidized and N-dealkylated metabolites (2.5%) were also detected. Similar metabolite profiles were also found in the 150 mg/kg BID dose group.

At 4 hours post-dosing on day 4, the average percentages of BCL6 levels in WBCs of the groups at 50, 150 and 500 mg/kg BID BMS-986458, relative to vehicle group average, were 102.6%, 90.8% and 91.3%, respectively, indicating no significant decrease in BCL6 at ∈500 mg/kg BID. In BMS-986458 dosed rats, basal BCL6 expression in circulating WBCs is low and may interfere with the ability to detect decreased.

There was no moribund or dead or impact on clinical observations, body weight, food consumption, organ weight, gross pathology or histopathology associated with BMS-986458.

At > 50 mg/kg BID, BMS-986458 related findings include:

At doses greater than or equal to 150 mg/kg BID, minimal reduction in the total mature red blood cell parameters (RBC, hematocrit [ HCT ], hemoglobin [ HGB ]) correlates with minimal to mild dose-related reduction in reticulocyte count, without microscopic-related abnormalities

Minimal usual dose-related increase in total WBC lymphocytes, monocytes (at > 150 mg/kg BID), basophils and/or large undyed cell counts, minimal decrease in non-histologically related abnormal triglycerides, minimal to mild decrease in total bile acid concentration, and minimal dose-related increase in cholesterol concentration

Findings of triglycerides and cholesterol are considered to be potentially pharmacologically mediated in view of reports (knockout mouse studies) ^65,66 describing the emerging role of BCL6 in lipid metabolism and/or also potentially due to reduced bile acid synthesis ⁶⁷

At a high dose of 500 mg/kg BID, a slight prolongation of BMS-986458 was observed in relation to Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT), with no clinically relevant abnormalities.

In summary, male rats were clinically well tolerated by BMS-986458 within 5 days at an oral dose of 500 mg/kg BID or less. Based on the absence of dose limiting toxicity, the MTD was considered to be 500 mg/kg BID (BMS-986458 mean AUC [0-24h ] of 149 μg.h/mL).

4.3.3.2 Exploratory toxicological kinetics and tolerability studies in female rats that were orally administered BMS-986458 (CC-2021676) twice daily for 10 consecutive days. In the exploratory toxicodey and tolerability study ⁴³, BMS-986458 was administered to female rats (6 per treatment group) at doses of 0, 50, 150 and 300 mg/kg BID (6 hour dosing interval) for 10 days. Evaluation criteria included toxicology, BCL6 protein measurement, survival, body weight, food consumption, clinical observations, clinical pathology, histopathology, and hepatic CYP transcription profile.

For BMS-986458 total drug concentration or each individual diastereomer concentration, lower dose-proportional exposures were observed. A60:40 ratio of individual isoforms (BMT-648592: BMT-648599) was detected in plasma, favoring BMT-648592. The pharmacokinetic data are summarized below.

TABLE B3-4.3.2.2-1 summary of toxicology-average-exploratory toxicology and tolerability study in female rats orally administered BMS-986458 (CC-2021676) twice daily for 10 consecutive days

BCL6 protein in spleen, ileum and liver was measured about 18 hours after the final dose. At high doses, there is a potential rebound of BCL6 protein expression in spleen, ileum and liver, ranging from about 1.3 to 1.7× (p≤0.06).

At all doses, BMS-986458 was well tolerated with no changes in body weight, clinical observations, or serum chemistry. There was a dose-related increase in WBC at 50, 150 and 300 mg/kg BID, respectively, with fold changes of 1.54, 1.85 and 2.95. In all dose groups, no adverse histopathological findings were present in liver, spleen, mesenteric lymph nodes and ileum.

The group mean increase in absolute liver weight was not statistically significant (to 13%) at 150 and 300 mg/kg BID compared to vehicle controls, with no histopathological related abnormalities.

Lymphocytes were increased 3-fold and monocytes were increased 3.7-fold at 300 mg/kg BID compared to control. Also at this dose level, all animals showed minimal proliferation of the myeloid lineage and/or depletion of the marginal zone in the spleen, which is considered pharmacologically relevant.

Transcript expression of CYP2B2, CYP3A23 and CYP4A2 in liver samples is measured and Ehhadh (enoyl-CoA, hydratase/3-hydroxyacyl-CoA dehydrogenase) encoding a bifunctional enzyme important in the peroxisome beta-oxidation pathway is encoded. There was a statistically significant decrease (about 0.5×, 0.2×, and 0.25×) in CYP2B2 transcriptional expression and a statistically significant increase (about 3.5×,7×, and 14×) in CYP3a23 transcriptional expression at 50, 150, and 300 mg/kg BID, respectively.

Based on the findings in this study, BMS-986458 had an MTD greater than or equal to 300 mg/kg BID.

4.3.2.3 Two week oral BID exploratory study in female rats. In the exploratory two-week toxicity study ⁴⁴, BMS-986458 was administered to a group of 6 female rats by oral gavage BID (6±0.5 hours apart) for 13 days with a total daily dose of 0, 100, 300 or 600 mg/kg/day (0, 50, 150 or 300 mg/kg BID). On day 14, a single dose of 0, 50, 150 or 300 mg/kg was administered prior to necropsy for timed collection of genotoxicological samples. All BMS-986458 doses were administered either at 10 mL/kg (20 total mL/kg daily) BID in vehicle consisting of 10% Captisol (sodium sulfobutylether β -cyclodextrin) and 90% 25 mM citrate buffer pH 3±0.1, or once at 10 mL/kg the last day. For genotoxicity assessment, positive control groups of 6 female rats were administered 10 mg/kg/day Cyclophosphamide (CP) by oral gavage on days 11 and 12, and 200 mg/kg/day Ethyl Methanesulfonate (EMS) on day 14, both at 10 mL/kg. Endpoints include survival, toxicology, clinical observations, body weight, clinical pathology (hematology, coagulation and serum chemistry evaluations), immunological (plasma cell phenotyping and peripheral blood lymphocyte phenotyping) evaluations, organ weight, gross and microscopic pathology analyses, BCL6 protein evaluations in spleen and ileum, micronucleus evaluations, and comet assay evaluations. Scheduled necropsies were performed 2 weeks after BID dosing.

Evaluation of BMT-986458 and its diastereomers BMT-648592 and BMT-648599 demonstrated a dose-proportional increase in concentration at day 1 at 4 hours post-administration of BMT-648592, BMT-648599, and thus BMS-986458. On day 13, the BMT-648592 and BMT-648599 concentrations at 4 hours post-dose were 4-2 times the 1 st day. After repeated dosing, the exposure to BMT-648592 and BMT-648599, and thus BMS-986458 (AUC [0-24h ] and Cmax), was less than dose proportionality. BMT-648592 and BMT-648599 were biased toward BMT-648592 at a ratio of about 2:1 in terms of day 13 exposure (AUC) and 4 hours after day 1 dosing.

A summary of the toxicology of BMS-986458, BMT-648592, and BMT-648599 is presented in Table 4.3.2.3-1 below.

Table B3-4.3.2.3-1 toxicity kinetics summary-average-two week oral BID exploratory toxicity study in female rats

At all doses, BMS-986458 was well tolerated and had no effect on survival, clinical observations, body weight, clotting, micronucleus assessment and comet assay.

At > 50 mg/kg BID, reduced pharmacodynamic changes in BCL6 protein (27% to 44% BCL6 protein degradation) were exhibited in the spleen.

BMS-986458 related findings are generally dose dependent and include:

at 50 mg/kg BID or more:

A minimal increase in reticulocyte count and a corresponding increase in the width of the distribution of red blood cells indicate an appropriate bone marrow response to a minimal non-dose related decrease in mature RBC parameters (RBC, HGB and HCT)

Minimal to mild increases in total WBC and constitutive lymphocyte, monocyte, basophil count and/or large undyed cell count

Increase in peripheral plasma cells (4×, compared to control group)

Non-dose related increases in total bilirubin levels to a minimum to a slight extent, without histologically related abnormalities

Minimal to mild non-dose related increase in phosphorus concentration

Minimal to mild increase in cholesterol

The increase in liver (absolute and/or relative to body weight) weight was associated with an increased liver size in 1 rat each at 300 and 600 mg/kg/day, a minimal increase in hepatocyte mitosis in 1 rat at 300 mg/kg/day, and minimal hepatocyte hypertrophy at all dose levels.

At 150 mg/kg BID or more:

Minimal increase in platelet count

A very small increase in globulin concentration, along with an increase in white blood cell count seen at > 100 mg/kg/day, indicates the possible presence of an immune/inflammatory response without histologically related abnormalities

T cells, helper T cells, cytotoxic T cells, B cells and NK cells were increased (up to 2X) compared to the control

At 300 mg/kg BID:

minimal increase in glucose concentration, combined with minimal increase in cholesterol at a dose of ≡100 mg/kg/day, indicates a slight effect on metabolic parameters

In summary, female rats were clinically well tolerated by BMS-986458 within 13 days when administered by oral gavage at a total daily dose of 150 mg/kg BID (single dose of 150 mg/kg on the day of necropsy). The main target organ at a dose of 50 mg/kg BID (average AUC on day 13. Gtoreq.216. Mu.g.h/mL) is liver (minimal hepatocyte hypertrophy and increased mitosis). Based on the finding that the MTD was found to be at least as high as 300 mg/kg BID (average AUC on day 13. Ltoreq.335. Mu.g.h/mL).

4.3.2.4 One month oral toxicity study with 2 week recovery period in rats. In a critical one month toxicity study ⁴⁵, BMS-986458 was administered to a group of 15 rats/sex at a dose of 0 (reference), 50, 150 or 500 mg/kg BID (8.+ -. 0.5 hours apart; total daily dose of 0, 100, 300 or 1000 mg/kg/day). High grade fine particulate hydroxypropyl methylcellulose acetate succinate (HPMC-AS-HF) at a concentration (149.04 mg/mL) equivalent to that of HPMC-AS-HF in BMS-986458 high dose (group 4) formulations was administered to the control group. All doses were administered at10 mL/kg in a vehicle/vehicle consisting of 0.25% w/w Methocel E4M in ultra pure water.

Evaluation criteria included survival, TK parameters, clinical observations, physical examination, body weight, food consumption, ophthalmic examination, clinical pathology assessment (hematology, coagulation, clinical chemistry and urinalysis), cytokine assessment, BCL6 protein levels in circulating WBCs analyzed by LC-MS, peripheral blood immunophenotyping (total T lymphocytes, helper T lymphocytes and cytotoxic T lymphocytes, B lymphocytes, NK cells), T cell dependent antibody response (TDAR) analysis, organ weight, and general and microscopic pathology. Scheduled necropsies were performed 1 month after dosing (10/sex/group, all groups) and after a 2 week recovery period (5/sex/group, all groups).

On day 28, the average systemic exposure (AUC [0-24h ]) of BMS-986458 and its diastereomers BMT-648592 and BMT-648599 was generally less than dose-proportional increase in the dose range of 50 to 500 mg/kg BID (100 to 1000 mg/kg/day) and without significant sex differences, given the large inter-individual variability. For all analytes, the average AUC (0-24 h) value at day 28 was slightly lower (0.4 to 0.8×) at day 1. On days 1 and 28, the average systemic exposure (AUC [0-24h ]) of BMT-648592 was about 2.5 to 3.3× of the other diastereomer BMT-648599. The diastereomeric ratio was generally similar for all doses and did not change substantially after repeated dosing.

The summary of the toxicology of BMS-986458 and its diastereomers is presented in the following table.

Tables B3-4.3.2.4-1 summary of toxicology-average sex pool-one month oral toxicity study with 2 week recovery period in rats

BCL6 protein was quantified in all leukocyte samples tested. BCL6 levels in circulating WBCs were not significantly regulated at any dose level relative to the control. In BMS-986458 dosed rats, basal BCL6 expression in circulating WBCs is low and may interfere with the ability to detect decreased. BCL6 is an immune function modulator, and the immunodiscovery results described in the following sections (including immune cell counts, cytokine levels, and changes in antibody response to KLH) are consistent with pharmacological activity downstream of BCL6 degradation. ¹⁰'¹¹

BMS-986458 was clinically well tolerated at all doses and had no BMS-986458-related death or effect on body weight, food consumption, clinical observations, physical examination, ophthalmology, urinalysis, or immunophenotyping endpoints. No BMS-986458-related adverse findings were noted at any dose level. Although BMS-986458 related, the following changes are not considered undesirable because they have minimal severity and/or lack toxicological significance.

At doses > 50 mg/kg BID, BMS-986458 related findings include:

Slight increase in globulin and slight decrease in albumin to globulin ratio (A/G) at > 150 mg/kg BID

Dose-independent mild increases in mean urea nitrogen in males and females alone at > 150 mg/kg BID, and no microscope-related abnormalities

A slight decrease in total bile acid in females and in males at 500 mg/kg BID, and a minimal increase in cholesterol in females alone at > 150 mg/kg. These changes are potentially pharmacologically mediated in view of reports describing the emerging role of BCL6 in lipid metabolism and/or due to reduced bile acid synthesis ^65,66,67

Average monocytes, lymphocytes (. Gtoreq.150 mg/kg BID), WBC (. Gtoreq.150 mg/kg BID), large undyed cells (male, at 500 mg/kg BID) and a slight increase in platelet count (. Gtoreq.150 mg/kg BID)

Mild or moderate increase in extramedullary hematopoiesis in males and in females at > 150 mg/kg, correlated with an increase in spleen weight (average absolute and relative values [ relative to body and brain ]; up to 31%) at > 150 mg/kg BID, a very small to mild increase in distribution width of erythrocytes and reticulocyte count at 50 mg/kg BID only, and a very small decrease in average erythrocyte parameters (RBC, HGB and/or HCT) at > 150 mg/kg BID

Increases in liver weight in females (average absolute and relative values [ relative to body and brain ]; up to 31%) in males and at > 150 mg/kg BID, correlated with minimal hypertrophy of lobular central hepatocytes in livers at 500 mg/kg BID

Minimal hypertrophy of the thyroid follicular epithelium, associated with increases in thyroid weight (mean absolute and relative values [ relative to body and brain ]; up to 30%), and is believed to be secondary to findings in the liver

Mild or moderate decrease in splenic marginal zone lymphoid cells

Reduction of serum TNFα (to 0.2×), IL-6 (to 0.1×), IFNγ (to 0.1×) and IL-2 (to 0.3×) levels in females and males at 150 mg/kg BID on day 28, and reduction of serum IL-1β (to 0.6×) in males at 500 mg/kg BID on day 28

At > 150 mg/kg BID, BMS-986458 related findings include:

Minimal reduction (shortening) of PT in females and in males at 500 mg/kg BID

Increases in adrenal weight (mean absolute and relative values [ relative to body and brain ]; up to 38%) in females and in males at 500 mg/kg BID, are associated with minimal hypertrophy of the fascicular bands of the adrenal cortex in females only at 500 mg/kg BID

At 500 mg/kg BID, additional BMS-986458 related findings include:

Minimal increase in phosphorus in females

Minimal increase in potassium in males

Reduced levels of anti-KLH IgM (day 8) and anti-KLH IgG (days 15 and 29) only after KLH immunization in females (to 0.4X in all time points and Ig isotypes)

At the end of the 2 week recovery period, all BMS-986458 related findings were completely reversible except for a slight non-dose related increase in urea nitrogen in females at > 50 mg/kg BID and in males at > 150 mg/kg BID.

In summary, rats were well tolerated BMS-986458 at BID oral doses of < 500 mg/kg BID over 1 month. No BMS-986458 related adverse findings were present at any dose. At the end of the administration, BMS-986458 related non-adverse findings included changes in serum chemistry parameters (potassium, phosphorus, BUN), RBC parameter reduction and regeneration response, thrombocytopenia, immune parameter (cell count, cytokines, antibody response to KLH) changes, and microscopic findings of adrenal glands, thyroid glands, liver and spleen. All BMS-986458 related changes were completely reversible except for a mild, non-dose related increase in urea nitrogen. The level at which no adverse effect was observed (NOAEL) was considered to be ≡500 mg/kg BID (average gender combined day 28 AUC [0-24h ] = 229 μg.h/mL of BMS-986458). No severe toxicity was observed and, therefore, STD10 was considered to be > 500 mg/kg BID, the highest dose tested.

4.3.2.5 Tolerability and toxicodel studies in dogs given twice daily for 5 consecutive days. In five-day toxicology and tolerability studies ⁴⁶, BMS-986458 was administered to a group of 3 dogs (mixed sex/group) by oral gavage at doses of 0 (vehicle), 7.5, 15 or 30 mg/kg BID (6 hours apart; 15, 30 or 60 mg/kg/day total). All doses were administered at 3 mL/kg in vehicle/carrier consisting of 10% Captisol in 25 mM citrate buffer (pH 3). Evaluation criteria included toxicology, BCL6 protein assessment in leukocytes by proteomics, potential changes in off-target protein levels in leukocytes by proteomics, survival, body weight, food consumption, clinical observations, FOB evaluation, and clinical pathology (serum chemistry and hematology) evaluation.

Dose proportionality was observed between the dose groups of 7.5 mg/kg BID to 30 mg/kg BID for individual diastereomeric concentrations, and between 15 mg/kg BID and 30 mg/kg BID for day 1. The plasma concentrations of the individual isoforms of BMS-986458 (BMT-648592 and BMT-648599) have a tendency to favor BMT-648592 at a ratio of 2:1. The summary of the toxicology of BMS-986458 and 2 isomers is presented in the following table.

TABLE B3-4.3.2.5-1 summary of toxicology-average-tolerability and toxicology studies in dogs given twice daily for 5 consecutive days

The dose-independent BMS-986458-related degradation of BCL6 in peripheral blood leukocytes was noted at about 4 hours and 24 hours after dosing on days 1 and 4 morning, indicating sustained BCL6 degradation (> 80% reduction compared to control) throughout the dosing period. Exemplary CELMoD molecular gum substrates (e.g., IKZF1/3, GSPT, CK 1. Alpha., HBS 1L) were unaffected. BCL6 is selectively down-regulated at the early collection point. A modest change in leukocyte proteome was observed in subsequent samples collected after day 4 dosing.

BMS-986458 associated body weight or clinical chemistry changes were not noted at any dose level.

At doses ≡7.5 mg/kg BID, BMS-986458 started to cause sporadic stool changes (including unshaped or liquid stool or no stool discharge) at day 1 and became most common by day 5, with some dogs experiencing stool changes during the post-dosing period until day 8. On day 4 and/or day 5, fecal samples from all dogs treated with BMS-986458 were hematochezia positive. A decrease in food consumption (up to 50% decrease) was observed from day 3. In addition, all dogs treated with BMS-986458 exhibited elevated WBCs and neutrophils by day 5 and continued until day 15.

In summary, BMS-986458 was orally administered to dogs at 7.5, 15 and 30 mg/kg BID for 5 days to achieve dose-proportional exposure and sustained BCL6 degradation in leukocytes > 80% throughout the study. At all doses, BMS-986458 caused fecal changes, including hematochezia, and had evidence of recovery during the post-dosing period.

4.3.2.6 Five days in female dogs. In a 5-day exploratory toxicity study in a group of 2 female dogs ²⁸, control preparations were orally administered at a dose volume of 5 mL/kg BID (8±0.5 hours apart) for 5 days (1 administration on day 5 only) as an introduction period to assess tolerance of control preparations. BMS-986458 was orally administered daily to the dogs for 5 days (days 15-19) at doses of 5, 15 or 50 mg/kg BID (8+ -0.5 hours apart; total daily dose of 10, 30 or 100 mg/kg/day) after a period of 9 days of no dosing (days 6-14), except on day 19 when the dogs received only 1 dose. All doses were administered at 5 mL/kg BID in vehicle/vehicle consisting of 0.25% Methocel E4M in water. Endpoints include survival, toxicology, metabolite profiling, clinical observations, body weight, food consumption, clinical pathology and exploratory FOB test evaluations, targeted protein (BCL 6) expression in leukocytes, organ weight, and gross and microscopic pathology analyses. Scheduled necropsies were performed after dosing at the end of the 5 day dosing period.

During the 5-day control introduction period, no findings related to the control were noted.

After repeated dosing, the average systemic exposure (AUC [0-24h ]) of BMS-986458 and its diastereomers BMT-648592 and BMT-648599 is generally increased from 5 to 15 mg/kg BID more than dose proportionality and from 15 to 50 mg/kg BID approximately dose proportionality. The summary of the toxicology of BMS-986458, BMT-648599, and BMT-648592 is presented in the following table.

Table B3-4.3.2.6-1 summary of toxicology-average-five day oral exploratory study in female dogs

The metabolite profile of the dog plasma samples showed no change in BMS-986458 as the primary drug species in circulation, accounting for about 90% of the total circulating species in dog plasma, following doses of 15 or 50 mg/kg BID. The most abundant metabolites in dog plasma are formed via parental demethylation (M11) and oxidation (M9), accounting for 1.6% and 3.3% of total circulating species. Low levels of oxidative, dehydrogenated, dealkylated and hydrolytic metabolites (< 0.4%) were also detected.

BCL6 concentrations in 5 white blood cell samples from 6 dogs were below the limit of quantitation (LOQ) 4 hours after the first dose at BMS-986458 fourth day, except slightly above LOQ at low doses of 5 mg/kg BID. This finding indicates BCL6 degradation at all dose levels, as samples from untreated dogs were higher than LOQ in another study. ⁴⁷

There were no BMS-986458 related moribund/dead or effects on feeding behavior, organ weight or general pathology.

At > 15 mg/kg BID, BMS-986458 related findings include:

Minimal reduction in body weight compared to the beginning of the dosing period (compared to the beginning of the dosing period-0.15 kg)

FOB positive in 4 dogs with a dose of 15 mg/kg BID, no clinical observation or microscopic abnormalities, FOB+ pre-test at 1/5 (1 sample not submitted)

At a high dose of 50 mg/kg BID, BMS-986458 related findings include:

clinical observations of 1 dog on diluted feces on day 3 and on day 4

Minimal increase in monocyte count and fibrinogen concentration, indicative of inflammatory response, and no histopathologically related abnormalities

Minimal increase in cholesterol concentration

Minimal to moderate reduction of lymphoid cell mass in GALT of thymus, mandibular lymph nodes and stomach and colon in 1 female dog

In summary, female dogs were clinically well tolerated to BMS-986458 within 5 days at oral doses of 50 mg/kg BID or less. Based on the absence of dose limiting toxicity, the MTD was considered to be 50 mg/kg BID (average AUC [0-24h ] of BMS-986458 was 2,120 μg.h/mL).

4.3.2.7 One month oral toxicity study with 2 week recovery period in dogs. In a critical one month toxicity study ⁴⁷, BMS-986458 was dosed at 0 (vehicle), 5, 15, or 50 mg/kg BID (8.+ -. 0.5 hours apart; total daily dose of 0,10, 30, or 100 mg/kg/day) by BID oral gavage to groups of 5 dogs/sex in groups 1,3, and 4, and groups of 3 dogs/sex in group 2. All doses were administered at 5 mL/kg in a vehicle/vehicle consisting of 0.25% w/w Methocel E4M in ultra pure water. The control group was administered HPMC-AS-HF at a concentration (29.81 mg/mL) equivalent to that in the BMS-986458 high dose (group 4) formulation. Endpoints include survival, toxicology, clinical observations, body weight, food consumption, physical (including respiratory and nervous systems) and ophthalmic examinations, electrocardiography, clinical pathology assessment (hematology, coagulation, clinical chemistry, urinalysis and FOB), cytokine assessment, BCL6 protein expression in circulating WBCs obtained using LC-MS and BCL6 protein expression in lymphoid tissues obtained by immunohistochemistry, immunophenotyping (total T cells, helper T cells, cytotoxic T cells and B cells), TDAR analysis, organ weight, and gross and microscopic pathology analysis. Scheduled necropsy was performed 28 days after dosing (3/sex/group) and after a2 week recovery period (2/sex/group, groups 1,3 and 4).

After repeated dosing, BMS-986458 and the average systemic exposure of diastereomers BMT-648592 and BMT-648599 (AUC [0-24h ]) were generally increased from 5 to 15 mg/kg BID more than dose proportionality and from 15 to 50 mg/kg BID approximately dose proportionality, and given the large inter-individual variability, no significant sex differences were noted and no accumulation or loss of exposure. On days 1 and 28, the average systemic exposure (AUC [0-24h ]) of BMT-648592 was about 2.7 to 3.7× of the other diastereomer BMT-648599. The diastereomeric ratio was generally similar for all doses and did not change substantially after repeated dosing.

Table B3-4.3.2.7-1 summary of toxicology-average sex pool-one month oral (gavage) toxicity study with 2 week recovery period in dogs (Source: study DN22136 ⁴⁷)

BCL6 degradation was assessed by immunohistochemistry of lymph nodes and LC-MS analysis of circulating leukocytes. Immunohistochemical analysis of lymph nodes from control dogs showed robust staining of BCL6 in the hair center. BCL6 staining was negative and germinal centers were reduced or absent in all dose groups. After a 2 week recovery period, BCL6 staining at 15 mg/kg BID recovered, but 2 out of 4 dogs at 50 mg/kg BID remained negative for BCL 6. Consistent with immunohistochemical findings, significantly lower BCL6 levels were measured in circulating leukocytes at all dose levels, on days 1 and 28, as compared to controls.

BMS-986458 related deaths were noted at 50 mg/kg BID. From day 1, a significant decrease in food consumption was noted at 50 mg/kg BID, resulting in worsening of clinical conditions (weight loss, reduced activity, wasting status and/or localized tremors). All 50 mg/kg BID dogs were provided with nutrition fortification and the nutrition fortification improved the status of all dogs except 3, and these dogs were euthanized on day 7 or day 8. In 1 early dead, minimal neutrophil inflammation of the atrioventricular valve and right atrium of the heart was observed, but it was not considered an effect directly related to the test item, although it may be secondary to lymphoid changes. The cause of the clinical signs that require euthanasia to be performed on these 3 dogs could not be determined from macroscopic examination or microscopic examination, but death was considered BMS-986458-related.

At any dose, there was no BMS-986458-related effect on the eye, electrocardiogram, nervous system endpoint, respiratory rate, urinalysis, or cytokine endpoint. The following BMS-986458 related changes are not considered undesirable because they have minimal severity and/or lack toxicological significance.

At > 5 mg/kg BID, BMS-986458 related findings include:

Dose-dependent minimal to mild increases in mean WBC count, neutrophil count and lymphocyte count

The presence of neutrophils and 1+ cytoplasmic vacuoles in the 2+ or 3+ bands indicates an inflammatory response, an increase in total lymphocyte count associated with an increase in absolute counts of total T cells, helper T cells and B cells

Minimal to slight decrease in mean RBC count, HGB and HCT, which correlates with microscopic observations of slight decrease in mean reticulocyte count, increase in red cell distribution width at 50 mg/kg BID, and decrease in bone marrow erythroid cell mass at 15 and 50 mg/kg BID for females and 15 mg/kg BID for males alone

Minimal to slight increase in cholesterol

Average liver weight gain in males (average absolute and relative values [ relative to body and brain ]; up to 47%) at > 15 mg/kg BID, correlated with microscopic hepatocyte hypertrophy and hepatocyte rarefaction

On days 15, 22 and 29, KLH-specific IgG responses were reduced compared to vehicle

At > 15 mg/kg BID, BMS-986458 related findings include:

Lower average weight/weight loss noted throughout the dosing period (male-4.7% at 15 mg/kg BID and male and female-11% and-8% respectively at 50 mg/kg/dose BID) compared to their corresponding pre-test values on day-1

The lower average food consumption noted for males during week 1 at > 15 mg/kg/dose BID and females at 50 mg/kg/dose BID is alleviated by providing additional dietary supplements in the form of canned foods to the 50 mg/kg BID group from day 7/6 until the end of dosing

Minimal to slight increase in total bilirubin and minimal increase in glucose

At a high dose of 50 mg/kg BID, BMS-986458 related findings include:

Clinical observations of reduced activity, spinal herniation and/or wasting conditions noted in most surviving males and females, which correlate with average weight loss

Minimal to slight increase in fibrinogen

Minimal increase in average triglycerides

Reduction of mean albumin in males and related reduction of A/G ratio

Average kidney weight increase (average absolute and relative values [ relative to body and brain ]; up to 18%) without microscopic-related abnormalities

Thymic gland in 1 male and 1 female is small, associated with extremely small to moderate (female) and moderate or severe (male) decreases in lymphocyte cell mass

Minimal mixed cell inflammation of the leg tibial joint was observed in 1 female and all 3 early dead people

At the end of the 1 month recovery period, all BMS-986458 related findings were completely reversible, except for the following:

Minimal reduction of RBC counts, HGB and HCT, however, a regeneration reaction consisting of minimal to slight increase in distribution width of erythrocytes and reticulocyte counts at > 15 mg/kg BID, along with increase in bone marrow cell mass at > 15 mg/kg BID and rebound effects of extramedullary hematopoiesis of the spleen was observed

Cell number reduction of GALT lymphocytes in females at > 15 mg/kg BID

In summary, administration of BMS-986458 resulted in premature euthanasia of 2 males (day 8) and 1 female (day 7) in the 50 mg/kg BID group due to clinical signs associated with weight loss and reduced food consumption. The cause of the clinical signs that require euthanasia to be performed on these 3 dogs could not be determined from macroscopic examination or microscopic examination, but is considered to be related to BMS-986458. NOAEL and HNSTD were considered to be 15 mg/kg BID (average gender pooled BMS-986458 auc [0-24h ] =400 μg.h/mL at day 28) since no adverse findings or morbidity was observed at 15 mg/kg BID.

4.3.3 Genotoxicity

4.3.3.1 In vitro. In Salmonella typhimurium (S. Tyrmium) (TA 98, TA100 and TA 1537) BMS-986458 and its individual diastereomers BMT-648592 and BMT-648599 ^48,49,50 were not mutagenized in the case of no metabolic activation at 400, 481 and 481 μg/plate, respectively, or in the case of S9 metabolic activation at 800, 1050 and 1050 μg/plate, respectively, in the exploratory bacterial back mutation assay. Similarly, in the clear GLP-conforming bacterial back mutation assay ⁵¹ in salmonella typhimurium (TA 98, TA100, TA1535, TA 1537) and escherichia coli (e.coli) (WP 2 uvrA), BMS-986458 was not mutagenized at up to 313 μg/plate without metabolic activation or with metabolic activation (activated liver S9), respectively.

In the exploratory micronucleus assay ^68,69,70,71, BMS-986458 and the individual diastereomers BMT-648592 and BMT-648599 were positive for micronucleus formation in CHO and/or TK6 cells at > 1.7, 9 and 1.5 μg/mL, respectively, without S9 metabolic activation and at > 1.7, 59 and 4.5 μg/mL, respectively, with S9 activation. In the multi-flow assay ⁷¹ in TK6 cells, a BMS-986458 related increase in H2AX and p53 protein was observed at > 3.87 ug/mL, and an increase in polyploid was observed at > 88 ug/mL. These findings indicate that in vitro BMS-986458 has chromosome breakage at low concentrations and induction of aneuploidy at high concentrations.

4.3.3.2 In vivo

In vivo micronucleus and comet non-GLP studies in rats. BMS-986458 was administered to a group of 6 female rats by oral gavage BID (6.+ -. 0.5 hours apart) for 13 days as part of the exploratory toxicity study, with a total daily dose of 0, 100, 300 or 600 mg/kg/day (0, 50, 150 or 300 mg/kg BID). On day 14, a single dose of 0, 50, 150 or 300 mg/kg was administered prior to necropsy for timed collection of genotoxicological samples. Positive control groups of 6 female rats were administered 10 mg/kg/day of CP by oral gavage on days 11 and 12, and 200 mg/kg/day of EMS on day 14. Evaluation criteria included peripheral micronucleus assessment and comet determination in liver and duodenum. At all doses, BMS-986458 was well tolerated and had no effect on micronucleus assessment and comet assay (300 mg/kg BID AUC [0-24h ] = 335 μg.h/mL and Cmax = 32 μg/mL).

In vivo micronucleus and comet GLP studies in rats. BMS-986458 was administered to male rats (6/group) for 4 days ⁵³ by oral gavage at a dose of 0 (negative control), 50, 150 or 500 mg/kg BID (100, 300 or 1000 mg/kg/day) twice daily (BID, 3 hours apart). All doses were administered at 10 mL/kg in a vehicle consisting of 0.25% w/w Methocel E4M in ultrapure water. Evaluation criteria included survival, toxicology, clinical observations, body weight, percentage of reticulocytes in peripheral blood and frequency of micronucleated reticulocytes, and comet tail intensity (tail DNA%) in liver and duodenum. Scheduled necropsy was performed 2 hours (±30 minutes) after the last dose on day 4.

After repeated dosing, the average systemic exposure (AUC [0-21h ]) of BMS-986458 and its diastereoisomers BMT-648592 and BMT-648599, as measured after the first daily dose on day 3, generally increased less than dose proportionality over the dose range of 50 to 500 mg/kg BID. These toxicological results are summarized in the following table.

Table B3-4.3.3.2-1. Summary of the toxicology of BMS-986458 in Male rats-day 3

BMS-986458 up to 500 mg/kg BID was well tolerated when dosed for 4 days and had no effect on clinical observations, body weight and reticulocyte percentage (indicative of lack of hematopoietic cytotoxicity). At any dose level, no statistically significant increase in micronuclear reticulocyte percentage or percentage of comet intensity in liver and duodenum was observed compared to negative control. The positive controls cyclophosphamide and ethyl methanesulfonate were able to perform as expected, proving that the study was able to detect genotoxic substances.

In summary, when administered to male Sprague Dawley rats via oral gavage twice daily for four consecutive days at doses (maximum viable dose) up to 500 mg/kg BID (average day 3 AUC (0-21 h) =254 μg.h/mL) of BMS-986458, BMS-986458 did not induce chromosomal or mitotic spindle damage (as assessed in micronucleus assays) and showed no evidence of DNA damage in the liver or duodenum in vivo comet assays.

4.3.4 Carcinogenicity. No carcinogenicity study has been performed with BMS-986458.

4.3.5 Reproduction and developmental toxicity. No studies of reproductive or developmental toxicity have been performed with BMS-986458.

4.3.6 Local tolerance. No study of local tolerance has been performed with BMS-986458.

4.3.7 Other toxicity studies

4.3.7.1 Phototoxicity assay in Balb/C373 mouse fibroblasts (DN 22066). Because BMS-986458 absorbs light between 290 and 700 nm, in vitro testing ⁵⁴ of BMS-986458 was performed in a neutral red uptake assay in Balb/c 3T3 mouse fibroblasts. BMS-986458 exhibited phototoxic potential when tested in 1% dimethyl sulfoxide/Du's phosphate buffered saline up to its solubility limit of 17.8 μg/mL.

4.4 Safety pharmacology. The potential off-target pharmacology (binding activity and/or functional activity) of BMS-986458 and its individual diastereomers BMT-648592 and BMT-648599 against a broad set of receptors, ion channels, transporters and enzymes was evaluated, including the assessment of hERG (Potassium), cardiac calcium and cardiac sodium ion channel function. Additional in vitro evaluations of BMS-986458, BMT-648592, and BMT-648599 were performed in hiPSC-CM. In vivo safety pharmacology assessments of BMS-986458 included exploratory cardiovascular telemetry studies in awake rats and dogs and anesthetized rabbits, as well as cardiovascular, respiratory and/or neurological endpoints assessed in rats and dogs as part of a critical 1 month oral toxicity study.

4.4.1 In vitro off-target interaction assessment. BMS-986458 and/or BMT-648592 and BMT-648599 were tested in vitro for their activity ^55,56,57 on a broad spectrum of pharmacological receptors, transporters, ion channels, nuclear hormones and enzymes at a maximum concentration of less than or equal to 62.8. Mu.g/mL. BMT-648592 and BMT-648599 inhibited Phosphodiesterase (PDE) 3 and PDE4 at approximately comparable concentrations, wherein the IC50 of PDE3 is 3.7 and 1.9 μg/mL, respectively, and the IC50 of PDE4 is 1.8 and 2.7 μg/mL, respectively. No inhibition of PDE3 or PDE4 was observed when BMS-986458 was tested as a mixture. When screened for a panel of liver and gall transport proteins and enzymes, BMS-986458 inhibited BSEP, MRP2, and UGT1A1, and the respective IC50 values were 2.4, 4.6, and 0.094 μg/mL.

4.4.2 Cardiovascular assessment

4.4.2.1 In vitro heart ion channel assay. In patch clamp assays ^58,59,60, BMS-986458, BMT-648592 and BMT-648599 inhibited cardiac ion channels in vitro at approximately comparable concentrations, with hERG IC50 of 0.44, 0.52 and 0.16 μg/mL, respectively. BMT-648592 and BMT-648599 inhibited cardiac L-type calcium channels by 9.4% and 7.4%, respectively, at the highest test concentration of 0.628 μg/mL.

4.4.2.3 Cardiovascular studies in telemetry rats. In a single day cardiovascular study in telemetry rats ⁶², BMS-986458 was evaluated after oral administration to investigate the potential to induce changes in arterial blood pressure and heart rate. BMS-986458 was administered to each group of 5 to 6 male rats, each with a telemetry implant, by oral gavage at 0, 50, 150 or 300 mg/kg BID (6 hours between the two doses; total daily dose 0, 100, 300 or 600 mg/kg/day). Vehicle control animals (0 mg/kg) received the same volume of test article carrier (10% Captisol in 25 mM citrate buffer pH 3). Evaluation parameters were systolic pressure (SBP), diastolic pressure (DBP), mean arterial blood pressure (MAP), heart Rate (HR), core Body Temperature (CBT) and voluntary activity, which were continuously monitored 24 hours before and 24 hours after dosing. Male satellite rats (n=3) without implanted instruments received 0, 50, 150 or 300 mg/kg BID BMS-986458 to study the plasma concentrations of BMS-986458 and its diastereoisomers BMT-648592 and BMT-648599.

50 The exposure amounts of mg/kg BID to 300 mg/kg BID were lower than dose proportionality. The exposure to BMT-648592 and BMT-648599 had an estimated 1:1 ratio, with plasma concentrations with a slight insignificant trend toward BMT-648592. The toxicological data are summarized in Table 4.4.2.3-1.

Table B3-4.4.2.3-1 summary of toxicology-average oral single day BID dose study on telemetered Male rats cardiovascular study

At up to 300 mg/kg BID, there were no BMS-986458 related changes in blood pressure (MAP, SBP, DBP), heart rate, or core body temperature. At doses of 150 and 300 mg/kg BID, BMS-986458 reduced voluntary activity during the period of time when the animal was most active.

4.4.2.4 Electrophysiological study in anesthetized rabbits. In electrophysiological studies in anesthetized rabbits ⁶³, BMS-986458 was administered intravenously over 30 minutes to 3 anesthetized male rabbits at incremental doses of 0.33, 2, or 12 mg/kg (12 mg/mL). An additional 3 anesthetized male rabbits were given an infusion of vehicle as a control. Intracardiac His-beam and surface Electrocardiographies (ECGs) were recorded continuously before, during and after incremental dosing. BMS-986458 concentrations were determined at the end of each infusion and in plasma collected 10, 20 and 30 minutes after the highest dose infusion was completed.

The plasma concentrations of BMS-986458 increased in a dose-dependent manner, on average 1.6, 12.2, 45.8 μg/mL (2.6, 19.5 and 72.9 μM) following intravenous infusion at doses of 0.33, 2 and 12 mg/kg, respectively.

At a high dose of 12 mg/kg, BMS-986458 resulted in an increase in heart rate correction QT interval of 51.2 ms (QTcF) and 24.4 ms (QTcV). At 2 and 12 mg/kg, BMS-986458 resulted in a blood pressure decrease of-11.6% and-34.4%, and an increase in heart rate of 10.8% and 34.1%, which interfered with the interpretation of the QT interval findings. At any of the tested doses, the compounds had no significant effect on PR, QRS, AH or HV intervals.

In summary, intravenous administration of BMS-986458 to anesthetized rabbits at a high dose of 12 mg/kg (plasma concentration of 45.8 μg/mL) resulted in an increase in QTc. A decrease in blood pressure and an increase in heart rate were observed at doses of 2 and 12 mg/kg and interfered with the interpretation of QT interval.

4.4.2.5 Single day (BID) oral exploratory cardiovascular telemetry studies in dogs. In the exploratory cardiovascular study in telemetry dogs ⁶⁴, BMS-986458 was administered to 6 female beagle dogs equipped with telemetry devices at a BID oral dose of 30 mg/kg (60 mg/kg/day) at a dose volume of 6 mL/kg. Vehicle (10% Captisol/90% 25 mM citrate buffer pH 3±0) was administered to dogs serving as their own controls one day prior to BMS-986458 administration. Evaluation criteria (including heart rate, systemic arterial blood pressure (systolic, diastolic and mean), left ventricular pressure (systolic and end diastolic), parameters of left ventricular pressure origin (+dp/dt, -dP/dt), ECG parameters (P-wave duration, PR, RR, QRS and QT interval), physical activity and core body temperature) were telemetered and evaluated during the 1 hour period prior to treatment with vehicle or BMS-986458 and approximately 21 hours after treatment. Clinical observations and heart rate corrected QT intervals (QTcA, QT intervals normalized to 80 beats/min heart rate) were also evaluated. The heart rate, rhythm or morphology of the ECG waveform was analyzed for interesting changes at about 4 hours and 10 hours post-dose, and plasma samples were collected at 6 hours and 24 hours post-dose for analysis of BMS-986458's 2 diastereomers BMT-648592 and BMT-648599.

Plasma concentrations of BMT-648592 and BMT-648599, as determined from samples taken 6 hours and 24 hours post-administration of BMS-986458, were substantially similar between animals and ranged from 1.26 to 24.5 μg/mL in individual animals. The results of the mean BMT-648592 and BMT-648599 plasma concentrations are summarized in the following table.

TABLE B3-4.4.2.5-1 average BMT-648592 and BMT-648599 plasma levels-Single day (BID) oral exploratory cardiovascular telemetry study in dogs

Drug or vehicle related clinical signs observed after treatment included unformed feces after BMS-986458 administration, which resolved within 48 hours.

A short minimal reduction in systolic blood pressure (about 11%) was observed 6 to 12 hours after the second BID dose. A short minimal decrease (about 7%) in left ventricular systolic pressure was noted about 10 to 12 hours after the second BID dose. After the second BID dose, left ventricular negative dP/dt was reduced (about 10%) overnight. There were no interesting changes in heart rate, other hemodynamic endpoints, ECG endpoints, physical activity or core body temperature after BMS-986458 administration. There was no BMS-986458 related change in the morphology of the ECG waveform.

In summary, BMS-986458 resulted in a short minimal decrease in arterial systolic and left ventricular systolic pressures and negative dP/dt overnight after a second oral BID dose of 30 mg/kg (60 mg/kg/day) in dogs, which resulted in average BMS-986458, BMT-648592 and BMT-648599 plasma concentrations of 16.21, 11.46 and 4.75 μg/mL, respectively, 6 hours after the first dose and 58.4, 39.5 and 18.9 μg/mL, respectively, 24 hours after the first dose.

4.4.2.6 Cardiovascular evaluation in a1 month GLP toxicity study in dogs. ECG recordings were performed approximately 2 hours after the first daily dose during week 4 in a1 month GLP toxicity study ⁴⁷ in dogs. No BMS-986458-related ECG findings were observed at any dose level (. Ltoreq.50 mg/kg BID; cmax=86.6. Mu.g/mL).

4.4.2.7 Respiratory and neurological assessment in a 1 month toxicity study in dogs. Respiratory and neurological assessments were performed 2 hours after the first daily dose during week 4 in a 1 month toxicity study ⁴⁷ in dogs. At any dose level (. Ltoreq.50 mg/kg BID; cmax=86.6. Mu.g/mL), no findings associated with BMS-986458 were noted for any of the parameters.

7. Data summarization

7.1 And (5) summarizing safety. Because there is no clinical experience with BMS-986458, the notes and safety monitoring programs of the initial clinical trial are based on findings of non-clinical toxicology studies (section 4.3 and 4.4). A summary of these findings, its potential clinical safety implications, and the requirements for monitoring safety by organ systems are summarized below. The fold exposure shown below in this section was calculated from the predicted exposure (AUC and Cmax) at the 40 mg BID initial clinical dose. The Cmax fold has been corrected for free fraction using a flux method using predicted species differences in protein binding and by assuming 99% protein binding for in vitro assays.

Since BMS-986458 is an experimental agent, it is possible that unforeseen, unknown or unexpected adverse events may occur. The investigator should review all relevant information in IB and the specific protocol considered (including inclusion/exclusion criteria) to determine suitability of the subject for group entry and appropriate evaluation of the subject after randomization into the study.

7.1.1 Immunomodulation. In repeated dose toxicity studies in rats and dogs, immunofindings consistent with immunosuppression and pro-inflammation were observed. For potential immunosuppression, a reduction in IgG and/or IgM antibody responses to KLH was observed in rats at doses of > 500 mg/kg BID (AUC [0-24h ] = 229 μg.h/mL;152×) and in dogs at > 5 mg/kg BID (AUC [0-24h ] = 47.5 μg.h/mL;31×) in a1 month toxicity study. Minimal to severe reduction in the amount of lymphoid cells in thymus, GALT and lymph nodes was also noted in dogs at > 5 mg/kg BID, characterized by a reduction in the size and number of germinal centers. The effects on IgM and IgG production and central pathology of lymph node germinal centers are consistent with the published role of BCL6 as the primary regulator of antibody affinity maturation in germinal centers ¹⁰ ^, ¹¹. Other findings included mild to moderate decrease of splenic marginal zone lymphoid cells and decrease of cytokines IL-2, IL-6, TNFα and IFNγ at > 50 mg/kg BID (AUC [0-24h ] = 129 μg.h/mL;86×) at day 28 and IL-1β at 500 mg/kg BID only in 1 month rat toxicity studies ⁴⁵.

Proinflammatory findings were noted in rats and dogs. In the 1 month rat toxicity study ⁴⁵, a slight increase in monocytes and globulins was noted at a dose of >50 mg/kg BID, and an increase in total WBC counts, large unstained cells and lymphocytes was observed at a dose of > 150 mg/kg BID (AUC [0-24h ] = 179 μg.h/mL;119×). In the 2 week non-GLP rat toxicity study ⁴⁴, minimal increases in total T cells, cytotoxic T cells, B cells and NK cells were noted at > 150 mg/kg BID (AUC [0-24h ] = 282 μg.h/mL;187×), but at similar exposure there were no similar changes in the GLP-compliant 1 month rat study. Similarly, in 1 month dog toxicity studies ⁴⁷ to a slight increase in WBC count, neutrophils, lymphocytes, total T cells, helper T cells and B cells were observed at >5 mg/kg BID. At 50 mg/kg BID (AUC [0-24h ] = 1,290 μg.h/mL;855×), minimal to moderate mixed cell inflammation associated with BMS-986458 of the leg shin joint was noted in 4 dogs, and minimal neutrophil inflammation in the atrioventricular valve of 1 dog.

Based on pre-clinical findings, monitoring of appropriate hematological parameters (WBC counts and classifications, immunophenotyping), antibody levels (IgA, igG, and IgM), cytokines, and physical examination, and guidelines for potential adverse event management are included in the clinical regimen.

7.1.2 Hematology-red blood cells and platelets. BMS-986458-related decreases in RBC parameters (RBC counts, HGB, HCT) were noted in the repeat dose toxicity study in rats and dogs. In the 1 month rat and dog GLP toxicity study ⁴⁵ ^, ⁴⁷, reduced RBC parameters were observed at doses of > 150 mg/kg BID (AUC [0-24h ] = 179 μg.h/mL;119×) and > 5 mg/kg BID (AUC [0-24h ] = 47.5 μg.h/mL;31×), respectively. In both species, a decrease in RBC parameters is associated with increased regeneration of reticulocytes, increased distribution width of red blood cells, extramedullary hematopoiesis, and/or rebound of cell mass in the bone marrow, with RBC parameters partially to fully restored during a 2 week recovery period.

In a1 month rat toxicity study, a minimal BMS-986458-related increase in platelets was observed at doses of > 150 mg/kg BID. Minimal increases in platelet count and/or changes in clotting time were noted in non-GLP repeat dose toxicity studies in rats, with no consistent changes in clotting time (both prolongation and shortening observed) ⁴⁴ in each study. No changes in platelet count or clotting time were noted in the dog repeat dose toxicity study.

Based on pre-clinical findings, appropriate monitoring of hematology parameters (red blood cell count and index, hemoglobin, hematocrit, and platelet count) and coagulation parameters (PT, APTT, and fibrinogen) and guidelines for potential hematologic adverse event management are included in the clinical regimen.

7.1.3 Arrhythmia potential. BMS-986458, BMT-648592, and BMT-648599 inhibited cardiac ion channels in vitro at comparable concentrations, with BMS-986458 hERG IC50 of 0.44 μg/mL, equivalent to a 518 Xfree Cmax multiple ⁵⁸ relative to a clinical starting dose of 40 mg BID. In vitro studies of hiPSC-CM ⁶¹, BMS-986458 was noted to increase FPDc, arrhythmia signal and decrease conduction velocity at concentrations ≡0.19 μg/mL (224×), and increased beat frequency was noted at 1.9 μg/mL (2240×).

BMS-98458 showed arrhythmogenic potential in anesthetized rabbits, but did not show arrhythmogenic potential in awake dogs when tested at comparable exposure. In exploratory rabbit electrophysiology studies ⁶³, a BMS-986458 related increase in QTc interval (qtcf=51.2 ms; qtcv=24.4 ms) was noted at administration of 12 mg/kg by IV infusion (plasma concentration=45.8 μg/mL;539×), but an increase in heart rate (34%) and a decrease in mean blood pressure (-34%) interfered with the explanation. ECG findings were not noted in dogs at 30 mg/kg BID in non-GLP telemetry studies or in 1 month GLP toxicity studies (Cmax. Ltoreq.86.6. Mu.g/mL; 192X).

Based on preclinical data, QT prolongation is not expected to occur at doses tested in the first human (FIH) trial. However, to identify any arrhythmogenic effects, ECG monitoring will be performed in the FIH trial.

7.1.4 Gastrointestinal toxicity. BMS-986458 related changes in stool composition (unformed stool or liquid stool) were noted at comparable exposures in 5 day dog exploratory toxicity studies rather than in 1 month GLP-compliant toxicity studies ^28,46,47 at doses ∈7.5 mg/kg BID (AUC [0-24h ] = 366 μg.h/mL;243×), and this was correlated with FOB test positives. Macroscopic examination or microscopic pathology related abnormalities in the gastrointestinal tract were not noted and recovery of the results was found after discontinuation of dosing. The FOB test in the 1 month study could not be explained due to the high positive rate of interference before the test and in the control. No fecal findings or gastrointestinal pathology findings were observed in the 1 month rat toxicity study at any of the high doses up to 500 mg/kg BID.

Based on preclinical data, appropriate monitoring and management of gastrointestinal toxicity is included in clinical protocols.

7.1.5 And (5) evaluating reproduction risk. BMS-986458 does not degrade selected classical CELMoD new substrates such as Aiolos, CK1 alpha, ikaros, GSPT1 and SALL4. Thus, it is expected that BMS-986458 toxicity profile will differ from CELMoD, including potential reproductive toxicity. However, the molecular basis of CELMoD-mediated reproductive toxicity has not been fully elucidated. Since no reproductive toxicity assessment has been performed so far, and the mechanism of BMS-986458 is cerebellar protein dependent, BMS-986458 was treated as a putative selective developmental toxic substance. No pathological findings were observed in male or female reproductive tissues at any dose in GLP toxicity studies at 1 month in rats and dogs.

Based on this, an appropriate pregnancy prevention program including consultation, pregnancy test and pregnancy prevention measures must be formulated for the clinical trial.

7.1.6 Liver risk assessment. BMS-986458 was identified as an inhibitor of BSEP, MRP2 and UGT1A1, and the corresponding IC50 was 2.4, 4.6 and 0.094 μg/mL. However, inhibition risk was low in the FIH clinical dose range, and the margin (margin) of free Cmax relative to these IC50 was greater than 100× for the 40 mg BID starting dose. In the 1 month dog toxicity study, minimal to slight increases in total bilirubin were noted at doses ∈15 mg/kg BID (cmax=25.1 μg/mL;56×), and other liver enzymes were unchanged. Increased bilirubin may be secondary to UGT1A1 inhibition because the free Cmax (0.251 μg/mL, assuming 1% unbound) at 15 mg/kg BID is higher than UGT1A1 IC50 in vitro. In contrast to the potential effects of BSEP, MRP2 and UGT1A1 inhibition, a decrease in circulating bile acid was noted in A1 month rat study at > 50 mg/kg BID (Cmax=11. Mu.g/mL; 129X).

In 1 month rat and dog toxicity studies ⁴⁵ ^, ⁴⁷, liver findings were limited to liver weight increases associated with minimal to mild hepatocyte hypertrophy, a finding consistent with BMS-986458 as a CYP inducer. It was considered that secondary to liver changes and unlikely to be human-related, minimal hypertrophy associated with BMS-986458 of thyroid follicular epithelium was only noted in rats at doses ∈50 mg/kg BID (AUC [0-24h ] = 129 μg.h/mL;86×).

In GLP-compliant 1 month rat and dog toxicity studies, minimal to mild BMS-986458-related increases in cholesterol were noted at doses of > 150 mg/kg BID (AUC [0-24h ] = 179 μg.h/mL;119×) and 5 mg/kg BID (AUC [0-24h ] = 47.5 μg.h/mL;31×), respectively. These findings correspond to an increasing number of documents demonstrating the role of BCL6 in the regulation of liver lipid metabolism. ⁶⁵ ^, ⁶⁶

Based on preclinical data, drug-induced liver injury is not expected to occur at doses within the FIH clinical dose range. All participants in the FIH study will be monitored for proper liver function at the appropriate frequency, including the assessment of transaminases, bilirubin and alkaline phosphatase. Although secondary thyroid findings are considered unlikely to be human-related, if TSH is abnormal, clinical safety laboratory assessments will be performed to monitor TSH and free T4. Based on preclinical lipid profile changes, clinical safety laboratory evaluations will be performed to monitor total cholesterol, triglycerides, low density lipoproteins and high density lipoproteins.

7.1.7 Phototoxicity. BMS-986308 absorbs light between 290 and 700 nm. The phototoxic potential of BMS-986458 was positive in a GLP-compliant neutral red uptake phototoxicity assay ⁵⁴ in Balb/c 3T3 mouse fibroblasts. Follow-up confirmation of phototoxic potential in vivo studies has not been completed.

Based on current preclinical data, patients should be provided with consultation to minimize exposure to ultraviolet light, wear protective clothing and sunglasses when out in the sun, and use broad spectrum Ultraviolet (UV) a/UVB sunscreens.

7.2 A human administration range and a safety margin. Clinical initiation doses were selected based on findings from 1 month toxicity studies in rats and dogs and PD studies in the OCI-LY1 xenograft mouse model. In the critical 28-day toxicity study in rats, no adverse findings were present and NOAEL was ≡500 mg/kg BID. Thus, STD10 > 500 mg/kg BID was considered. In a critical 28 day dog toxicity study, a decrease in food consumption was noted at a high dose of 50 mg/kg BID, resulting in weight loss and poor clinical status, leading to subsequent humane euthanasia in 3 dogs at the early days of 7, and an unknown cause of clinical status decline. There was no adverse effect at the intermediate dose of 15 mg/kg BID, and this dose was considered NOAEL and HNSTD. Dogs were used as the most sensitive species, and after application of the 6 x safety factor, the Maximum Recommended Starting Dose (MRSD) of 83 mg BID was calculated using the method described in ICH S9 (body surface area BSA conversion and growth scale at different rates).

Pharmacokinetic/pharmacodynamic and efficacy studies in the OCI-LY1 xenograft mouse model indicate that complete BCL6 inhibition is required for 24 hours to achieve maximum tumor growth inhibition. In humans, BMS-986458 was predicted to have low clearance (3.6 mL/min/kg) and moderate Vss (2.08L/kg), with a corresponding terminal half-life of about 10.6 hours. Based on the predicted terminal half-life in humans, BID administration is expected to achieve complete BCL6 inhibition for 24 hours. In this xenograft model, an effective dose range of 10 to 80 mg BID is currently expected based on tumor growth inhibition.

The initial dose in CA1231000 was chosen to be 40 mg BID. Since this dose is in the middle of the predicted effective dose range and is less than 83 mg BID MRSD, it is expected to be both effective and safe. The highest clinical dose will be determined based on human tolerance. The predicted AUC and Cmax at the highest proposed clinical dose 640 mg BID were at least 9.5 x and 3.5 x lower than the predicted AUC and Cmax at NOAEL in the critical 1 month rat and dog study, respectively.

The fold exposure presented in table 7.2-1 was calculated using a comparison of systemic exposure (Cmax and AUC) and BSA.

7.3.1 Pregnancy. BMS-986458 does not degrade classical CELMoD new substrates such as Aiolos, CK1 alpha, ikaros, GSPT1 and SALL4. Thus, BMS-986458 toxicity profiles would be expected to be distinct from CELMoD and IMID drugs (including thalidomide), including potential reproductive toxicity. However, since reproductive toxicity assessment has not been performed so far, and the mechanism of BMS-986458 is cerebellar protein-dependent, BMS-986458 was treated as a putative selective developmental toxic substance. Therefore, pregnancy prevention procedures must be followed in clinical procedures.

There is no data on the administration of BMS-986458 in pregnant females or animals and therefore it should not be administered to pregnant subjects. In the corresponding clinical regimen, the requirements for contraception and pregnancy screening are fully detailed for male and female participants.

7.3.2 Drug-drug interaction potential. Drug-drug interactions (DDI) have not been studied in clinical studies of BMS-986458.

Reactive phenotyping studies of the human cytochrome P450 (CYP) subtype and human liver microsomes indicate that BMS-986458 is metabolized primarily by CYP3A4/5 and that CYP1A2, CYP2C8, CYP2C9 and CYP2C19 are less involved. Furthermore, BMS-986458 is an inhibitor of CYP3A 4. These data indicate that there is a potential DDI if BMS-986458 is co-administered with a strong inhibitor or inducer of CYP3A4, a sensitive CYP3A substrate, or a CYP3A substrate with a narrow therapeutic window. If the use of a strong CYP inhibitor or inducer is necessary, its risk and benefit should be discussed with the Sponsor's medical inspector (Sponsor's Medical Monitor) before use with research interventions.

Examples of strong CYP3A inhibitors or inducers include (but are not limited to):

CYP3A inhibitors, such as clarithromycin, indinavir, itraconazole, ketoconazole, nefazodone, nelfinavir, ritonavir, saquinavir, telithromycin, grapefruit and grapefruit juice

CYP3A inducer, carbamazepine, phenytoin, rifampicin, and san John's Wart (St. John's)

A detailed list of strong CYP3A inhibitors or inducers, sensitive CYP3A substrates, or CYP3A substrates with a narrow therapeutic window is available on the FDA website.

Based on in vitro data, BMS-986458 may lead to inhibition of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP) in the gut, thereby enhancing oral absorption and plasma exposure of P-gp and BCRP substrates. At higher doses, BMS-986458 may also have the potential to inhibit liver uptake transporters (OATP 1B1 and OATP1B 3) and phase II drug metabolizing enzymes (UGT 1A 1). In view of the potential for such interactions, the toxicity signs of subjects receiving P-gp substrates (e.g., dabigatran, digoxin), BCRP substrates (e.g., rosuvastatin, sulfasalazine), P-gp or BCRP inhibitors (e.g., erythromycin, cyclosporin a) or OATP1B1 and OATP1B3 substrates (e.g., statins) or UGT1A1 substrates (e.g., valproic acid, levothyroxine or phenytoin) should be closely monitored.

Reference to the literature

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³Preclinical Investigation of In Vitro and Ex Vivo Response to BCL6 Degrader BMS-986458 in Models of Diffuse Large B-Cell Lymphoma (OG00458). Bristol-Myers Squibb, 2023. DCN 930205191.

⁴Crump M, Neelapu SS, Farooq U, et al, Outcomes in refractory diffuse large B cell lymphoma: results from the international SCHOLAR-1 study [published erratum appears in: Blood 2018;131(5):587-8]. Blood 2017; 130:1800-8.

⁵Purdum A, Tieu R, Reddy SR, Broder MS. Direct Costs Associated with Relapsed Diffuse Large B cell Lymphoma Therapies. Oncologist. 2019; 24:1229-36.

⁶Vaidya R, Witzig TE. Prognostic factors for diffuse large B cell lymphoma in the R(X)CHOP era. Ann Oncol 2014; 25:2124-33.

⁷Nagle SJ, Woo K, Schuster SJ, et al. Outcomes of patients with relapsed/refractory diffuse large B cell lymphoma with progression of lymphoma after autologous stem cell transplantation in the rituximab era. Am J Hematol 2013; 88:890-4.

⁸Gisselbrecht C, Glass B, Mounier N, et al. Salvage regimens with autologous transplantation for relapsed large B cell lymphoma in the rituximab era. J Clin Oncol 2010; 28:4184-90.

⁹Epperla N, Badar T, Szabo A, et al. Postrelapse survival in diffuse large B cell lymphoma after therapy failure following autologous transplantation Blood Adv 2019;3:1661-9.

¹⁰Bunting KL, Melnick AM. New effector functions and regulatory mechanisms of BCL6 in normal and malignant lymphocytes. Curr Opin Immunol. 2013 Jun;25(3):339-46

¹¹Basso K, Dalla-Favera R. Roles of BCL6 in normal and transformed germinal center B cells. Immunol Rev. 2012 May;247(1):172-83.

¹²Béguelin W, Teater M, Gearhart MD, et al. EZH2 and BCL6 Cooperate to Assemble CBX8-BCOR Complex to Repress Bivalent Promoters, Mediate Germinal Center Formation and Lymphomagenesis. Cancer Cell 2016; 30:197-213.

¹³Schlager S, Salomon C, Olt S, et al. Inducible knock-out of BCL6 in lymphoma cells results in tumor stasis. Oncotarget 2020; 11:875-90.

¹⁴Nonclinical Study Report: CEL-PD1920. Antiproliferative Activity of CC-99282 In Vitro (PD1920). Celgene Corporation, 2018.

¹⁵BMS-986458 In Vivo Pharmacology: Anti-tumor Activity of BMS-986458 in In Vivo Diffuse Large B Cell Lymhoma (DLBCL) Cancer Models (Study OG00457). Bristol-Myers Squibb, 2023. Document Control No. 930205190.

¹⁶Evaluation of BMS-986458 BCL6 Ligand Directed Degrader in Normal Immune Cell Populations (OG0468). Bristol-Myers Squibb, 2023. DCN 930205282.

¹⁷Effect of CC2021676 and its Single Isomers on In Vitro Neutrophil Maturation (OG00449). Bristol-Myers Squibb, 2023. DCN 930205189.

¹⁸Method Validation Report LCMSD 1206.2 Project RYHP2. Quantitation of BMT-648592 and BMT-648599 in Rat Plasma via HPLC with MS/MS Detection. PPD Richmond, VA, 26 May 2023. Bristol-Myers Squibb Document Control No. 930203824.

¹⁹Method Validation Report LCMSF 1206.3 Project RYJC2. Quantitation of BMT-648592 and BMT-648599 in Dog Plasma via HPLC with MS/MS Detection. PPD Richmond, VA, 21 April 2023. Bristol-Myers Squibb Document Control No. 930203826.

²⁰Nonclinical evaluation of the pharmacokinetics and distribution of BMS-986458 (NCPK781). Bristol-Myers Squibb Company, 2022. Document Control No. 930206028.

²¹Pharmacokinetic evaluation of BMS-986458 after a single intravenous or oral dose to male rats (NCPK778). Bristol-Myers Squibb Company, 2023. Document Control No. 930204665.

²²Pharmacokinetic evaluation of BMS-986458 after a single intravenous or oral dose to male dogs (NCPK782). Bristol-Myers Squibb Company, 2022. Document Control No. 930204591.

²³Pharmacokinetic evaluation of BMS-986458 after a single intravenous or oral dose to male monkeys (NCPK783). Bristol-Myers Squibb Company, 2022. Document Control No. 930204518.

²⁴In vitro characterization of ADME and DDI properties of BMS-986458 (NCPK785). Bristol-Myers Squibb Company, 2023. Document Control No. 930201163.

²⁵In vitro plasma protein binding and blood-to-plasma partitioning evaluation of BMS-986458 (NCPK832). Bristol-Myers Squibb Company, 2022. Document Control No. 930206106.

²⁶BMS-986458: Exploratory single dose 48 hour pharmacokinetics, pharmacodynamics and tolerability study in rats (DT21042). Bristol-Myers Squibb Company; 2022. Document Control No 930188328.

²⁷Nonclinical evaluation of the metabolism and drug-drug interaction potential of BMS-986458 (NCPK780). Bristol-Myers Squibb Company, 2021. Document Control No. 930103172.

²⁸BMS-986458: Five-day oral exploratory toxicity study in female dogs (DN22114). Bristol-Myers Squibb Company; 2022. Document Control No 930199213.

²⁹BMS-986458: Five-day oral exploratory toxicity study in female rats (DN22115). Bristol-Myers Squibb Company; 2022. Document Control No 930205113.

³⁰In vitro assessment of CYP induction potential of BMS-986458 in primary human hepatocytes (NCPK802). Bristol-Myers Squibb Company, 2022. Document Control No. 930203855.

³¹In vitro Evaluation of six selected test article as substrates of the human MDR1 Efflux (ABC) transporter (SOLVO Study Number BMS-31/NCPK863). SOLVO Biotechnology a Charles River Company, 2023. Document Control No. 930202386.

³²Prediction of human pharmacokinetics of BMS-986458 (NCPK864). Bristol-Myers Squibb Company, 2023. Document Control No. 930206283.

³³Guidance for Industry, Bioanalytical Method Validation. U.S. Department of Health and Human Services (USHHS), Food and Drug Administration (FDA), Center for Drug Evaluation and Research (CDER) and Center for Veterinary Medicine (CVM). 2018.

³⁴Guidance for Industry, Guideline on bioanalytical method validation. European Medicines Agency (EMA), EMEA/CHMP/EWP/192217/2009 Rev. 1 Corr. 2**. 2011.

³⁵Hosea, N. A., Collard, W. T., Cole, S., et al. (2009). Prediction of human pharmacokinetics from preclinical information: comparative accuracy of quantitative prediction approaches. J Clin Pharmacol, 49(5), 513-533. doi:10.1177/0091270009333209.

³⁶Davies, B., & Morris, T. (1993). Physiological parameters in laboratory animals and humans. Pharm Res, 10(7), 1093-1095.

³⁷Identification of metabolites of BMS-986458 in hepatocytes across species (NCPK826). Bristol-Myers Squibb Company, 2023. Document Control No. 930205962.

³⁸Guidance for Industry, Clinical Drug Interaction Studies - Cytochrome P450 Enzyme- and Transporter-Mediated Drug Interactions. U.S. Department of Health and Human Services (USHHS), Food and Drug Administration (FDA), Center for Drug Evaluation and Research (CDER). 2020.

³⁹Guidance for Industry, Guideline on the investigation of drug interactions. European Medicines Agency (EMA), CPMP/EWP/560/95/Rev, 1 Corr.2**. 2012.

⁴⁰Oie, S., & Tozer, T. N. Effect of altered plasma protein binding on apparent volume of distribution. J Pharm Sci 1979; 68(9): 1203-1205.

⁴¹BMS-986458: Maximum tolerated dose and toxicokinetic assessment of dogs administered a single oral dose of BMS-986458 (CEL-009463, CC2021676) (Study DT21066); Report and Amendment 1. Bristol-Myers Squibb Company; 2022. Document Control No. 930187418.

⁴²BMS-986458: Five-day oral exploratory toxicity study in male rats (Study DN22115). Bristol-Myers Squibb Company; 2023. Document Control No. 930205113.

⁴³Exploratory toxicokinetic and tolerability study in female rats orally administered BMS-986458 (CC2021676) twice daily for 10 consecutive days (Study DT22011); Report and Amendment 1. Bristol-Myers Squibb Company; 2023. Document Control No. 930199752.

⁴⁴CC2021676: Two-week oral BID exploratory toxicity study in female rats (Study DN21129); Report and Amendment 1. Bristol-Myers Squibb Company; 2023. Document Control No. 930203162.

⁴⁵BMS-986458: A one-month oral toxicity study in rats with a 2-week recovery period (Study DN22135). Bristol-Myers Squibb Company; 2023. Document Control No. 930204855.

⁴⁶BMS-986458 (CC2021676): Tolerability and toxicokinetic study in dogs dosed twice daily for 5 consecutive days (Study DT21078); Report, Amendment 1, and Amendment 2. Bristol-Myers Squibb Company; 2023. Document Control No. 930186855.

⁴⁷BMS-986458: A 1-month oral (gavage) toxicity study in dogs with a 2-week recovery (Study DN22136). Bristol-Myers Squibb Company; 2023. Document Control No. 930204949.

⁴⁸CEL-009463: Exploratory Ames reverse-mutation study in Salmonella typhimurium (Study DN21064); Report and Amendment 01. Bristol-Myers Squibb Company; 2021. Document Control No. 930175635.

⁴⁹BMT-648592: Exploratory Ames reverse-mutation study in Salmonella typhimurium (Study DN21135). Bristol-Myers Squibb Company; 2022. Document Control No. 930183007.

⁵⁰BMT-648599: Exploratory Ames reverse-mutation study in Salmonella typhimurium (Study DN21133). Bristol-Myers Squibb Company; 2022. Document Control No. 930183026.

⁵¹BMS-986458: Ames reverse-mutation study in Salmonella typhimurium and Escherichia coli (Study DN22069). Bristol-Myers Squibb Company; 2023. Document Control No. 930204435.

⁵²CEL-009463: Exploratory genotoxic mode of action using the multiflow assay in TK6 Cells (Study DN21111). Bristol-Myers Squibb Company; study on-going.

⁵³BMS-986458: Four-day oral micronucleus study with comet evaluation in male rats (Study DN22070). Bristol-Myers Squibb Company; 2023. Document Control No. 930204851.

⁵⁴BMS-986458: Neutral red uptake phototoxicity assay in Balb/c 3T3 mouse fibroblasts (Study DN22066). Bristol-Myers Squibb Company; 2023. Document Control No. 930201045.

⁵⁵BMS-986458/CEL-009463: In vitro assessment of receptors, ion channels, monoamine transporters, and enzymes (Study LDO00020). Bristol-Myers Squibb Company; 2023. Document Control No.930201989.

⁵⁶BMS-986458 isomer BMT-648592: In vitro assessment of interactions with receptors, ion channels, and enzymes (Study LDO00021). Bristol-Myers Squibb Company; 2023. Document Control No. 930201984.

⁵⁷BMS-986458 isomer BMT-648599: In Vitro assessment of interactions with receptors, ion channels, and enzymes (Study LDO00022). Bristol-Myers Squibb Company; 2023. Document Control No. 930201983.

⁵⁸BMS-986458: Effects on cardiac hERG channels (Study DT23033). Bristol-Myers Squibb Company; 2023. Document Control No. 930202916.

⁵⁹BMT-648592: Effects on cardiac hERG channels (Study DT23035). Bristol-Myers Squibb Company; 2023. Document Control No. 930202931.

⁶⁰BMT-648599: Effects on cardiac ion channels (Study DT23034). Bristol-Myers Squibb Company; 2023. Document Control No. 930202919.

⁶¹BMS-986458 (CEL-009463), BMT-648592 and BMT-648599: Exploratory in vitro multi-electrode array (MEA) study in human induced pluripotent stem cell-derived cardiomyocytes (Study DT23044); Report and Amendment 1. Bristol-Myers Squibb Company; 2023. Document Control No. 930204098.

⁶²BMS-986458 (CC2021676): Oral single day BID-dose investigative cardiovascular study in telemeterized male rats (Study DT21068); Report and Amendment 1. Bristol-Myers Squibb Company; 2022. Document Control No. 930186261.

⁶³BMS-986458: Electrophysiological effects in anesthethized rabbits (Study DT21051). Bristol-Myers Squibb Company; 2022. Document Control No. 930183352.

⁶⁴BMS-986458: Single day (BID) oral exploratory cardiovascular telemetry study in dogs (Study DT21079); Report and Amendment 1. Bristol-Myers Squibb Company; 2021. Document Control No. 930179833.

⁶⁵LaPensee CR, Lin G, Dent AL, et al. Deficiency of the transcriptional repressor B cell lymphoma 6 (Bcl6) is accompanied by dysregulated lipid metabolism. PLos One 2014; Jun 3;9(6):e97090.

⁶⁶Chikada H, Ida K, Nishikawa Y, et al. Liver-specific knockout of B cell lymphoma 6 suppresses progression of non-alcoholic steatohepatitis in mice. Sci Rep 2020 Jun 16;10(1):9704.

⁶⁷Chiang JYL. Bile acid metabolism and signaling. Compr Physiol 2013; Jul 3(3): 1191-212.

⁶⁸CEL-009463: Exploratory in vitro micronucleus study in Chinese hamster ovary cells (Study DN21094). Bristol-Myers Squibb Company; study on-going.

⁶⁹BMT-648592: Exploratory in vitro micronucleus study in TK6 cells (Study DN21136). Bristol-Myers Squibb Company; study on-going.

⁷⁰BMT-648599: Exploratory in vitro micronucleus study in TK6 cells (Study DN21134). Bristol-Myers Squibb Company; study on-going.

⁷¹BMT-648592, BMT-648599: Exploratory in vitro micronucleus study in TK6 cells (Study DN22072). Bristol-Myers Squibb Company; 2023. Document Control No. 930205597.

Example B4. clinical protocol CA 1231000

This example provides additional details of the phase I study described in example B2, and the disclosure of clinical regimen CA 1231000 (regimen revision 01; date: 2023, 6, 15; revision date: 2023, 8, 23) is incorporated herein by reference in its entirety.

References to chapters, tables, references, and/or appendices in example B4 refer to other chapters, tables, references, and/or appendices in example B4.

1. Scheme summary

Abbreviations AE, adverse events, BOIN, bayes optimum interval, CAR, chimeric antigen

Receptors, D, day, DLBCL, diffuse large B-cell lymphoma, DLT, dose limiting toxicity, ECOG, eastern tumor co-group, FDG, fluorodeoxyglucose, FL, follicular lymphoma, MTD, maximum tolerated dose, NHL, non-hodgkin lymphoma, OBD, optimal biologic dose, ORR, total response rate, PK, pharmacokinetics, R/R, recurrent/refractory, RP2D, recommended phase 2 dosing, SAE, severe adverse events, SCT, stem cell transplantation.

^a The primary measured (estimand) and supplemental measured sensitivity estimators will be defined in the statistical analysis plan (where applicable).

2. Activity schedule

If multiple procedures are required at a single point in time, ECG may be acquired up to 15 minutes in advance, vital signs may be acquired up to 10 minutes in advance or later, and clinical laboratory samples may be acquired up to 5 minutes in advance relative to the nominal point in time, ensuring that PK samples can be collected on time.

3. Introduction to the invention

3.1 The principle is studied. BMS-986458 is a small molecule bifunctional cerebellar protein-dependent ligand-directed-degrading agent (LDD) that will be investigated for treating patients suffering from recurrent and/or refractory (R/R) non-Hodgkin's lymphoma (NHL). This phase 1, first human (FIH) clinical study of BMS-986458 will explore the safety, tolerability, PK, pharmacodynamics, and preliminary biological and clinical activity of BMS-986458 as a single agent and in combination with an anti-lymphoma agent in the R/R NHL context.

3.2 Background

3.2.1. Disease background. NHL (the most common hematological malignancy worldwide) is a heterogeneous group of B-cell and T-cell lymphomas, accounting for approximately 4% of all cancers in the united states in total. Although there are more than 40 major subtypes, the most common subtypes include indolent Follicular Lymphoma (FL) and aggressive DLBCL.

3.2.1.1 Diffuse large B cell lymphomas. DLBCL is the most common subtype of NHL, accounting for 26% of all diagnosed NHL neoplasm cases, and it was estimated that 29,105 new cases ¹ occurred in the united states in 2020. The standard of care (SOC) currently used for these patients is a combination of CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) with rituximab (R-CHOP), however, about 10% to 15% of patients suffer from primary refractory disease, and 20% to 25% will eventually relapse after Complete Remission (CR) is reached, typically relapse ² in the first 2 years. The outcome of these patients remains poor, especially if the median total survival (OS) of refractory disease is about 6 months. ³

For many years, SOC for two-wire R/R DLBCL was high dose chemotherapy followed by autologous Stem Cell Transplantation (SCT). In the last few years, CAR-T cell therapy has been shown to cure most patients with R/R DLBCL (CR rates ranging between 40% and 58%) and has been approved for refractory disease or relapse ^4,5,6 within 12 months of first-line chemo-immunotherapy. DLBCL remains an incurable disease in recurrent or refractory patients who have received 2 or more normals of therapy (including SCT or CAR-T cell therapy) or are not candidates or are unable to obtain potential curative therapies due to advanced age or poor physical performance, an indication of which is clinically tested.

3.2.1.2 Follicular lymphoma. Follicular Lymphoma (FL) is the most common indolent lymphoma originating from germinal center B cells.

When therapy is indicated, the combination of anti-CD 20 immunotherapy with or without chemotherapy is the standard first line or subsequent line selection of late stage FL. About 20% of patients develop early relapse and disease progression within 24 months (POD 24), and these patients have poor outcome ⁷. It is not possible to identify these patients in advance and more effective treatment of these patients is required.

There are no standard treatments or sequences of treatments for R/R FL, and various methods include cytotoxic agents, anti-CD 20 antibodies, radiation therapy, radioimmunotherapy, and autologous and allogeneic transplantation. Chemotherapy regimens other than first-line therapy (bendamustine and rituximab [ BR ]; R-CHOP; or rituximab plus cyclophosphamide, vincristine, and prednisone [ R-CVP ]) are options in the context of relapse. Additional agents for multiple relapsed patients include lenalidomide, PBK inhibitors, tazemetostat, bispecific T cell conjugates (Mo Tuozhu mab), and CAR-T cell therapies. In addition to commercially approved targeted therapies for FL, there are a number of emerging agents targeting FL biology, including antibody-mug conjugates (ADCs), monoclonal or bispecific antibodies, checkpoint-based therapies, and novel combination strategies.

There is a highly unmet medical need to develop new treatments for FL patients that do not respond to standard therapies and for which treatment options for disease remission have been exhausted.

3.2.2. BMS-986458.BMS-986458 is a mixture of small molecules of diastereomers BMT-648592 and BMT-648599 bifunctional cerebellar protein-dependent LDD. LDD induces target protein degradation by recruitment of cerebellar protein (CRBN) E3 ubiquitin ligase, ubiquitination, and subsequent proteolytic processing by the 26S proteasome. Downstream effects of degradation include cell cycle arrest and apoptosis of tumor B cells and immunomodulation of T cells.

The in vitro sensitivity of tumor B cells to LDD is closely related to the expression level. Post-treatment transcriptomic responses and baseline genomics are used to define sensitivity profiles that incorporate posttranslational modulators and mechanical drivers of expression and LDD induction. High risk patient split clusters identified by lymphoma genome project (Lymphoma Genome Project) showed significant enrichment of LDD sensitivity characteristics.

BMS-986458 was evaluated in several in vitro NHL models and showed rapid and potent antiproliferative and pro-apoptotic tumor cell killing.

BMS-986458 exhibits potent and selective degradation, in vitro/in vivo efficacy in xenograft (CDX) models of various cell line origin, favorable PK/pharmacodynamic and toxicological characteristics in non-clinical safety assessment to enable assays in humans. A detailed description of the chemistry, pharmacology, efficacy, and safety of BMS-986458 is provided in the manual of researchers. 18

3.2.2.1 Non-clinical pharmacology. BMS-986458 in vivo pharmacology. BMS986458 as a single agent was evaluated in 2 DLBCL CDX models, where treatment showed a tumor volume reduction between 74% and 84% after oral administration at the highest dose. In all studies, this anti-tumor effect was achieved after greater than 98% degradation in tumor tissue, which was maintained throughout the administration period.

3.2.2. Non-clinical pharmacokinetics. In vitro and in vivo studies have been performed to characterize the absorption, pharmacokinetics, distribution, metabolism, and drug interaction potential of BMS-986458.

Following oral administration, metabolism of BMS-986458 was evaluated in human hepatocytes, as well as hepatocytes from preclinical species, and in plasma samples from rats and dogs. The primary metabolic pathways of BMS-986458 in the species tested include demethylation, oxidation, followed by glucuronidation and hydrolysis.

3.2.2.3 Non-clinical toxicology. The non-clinical toxicology of BMS-986458 (a mixture of diastereomers BMT-648592 and BMT-648599) was evaluated in a series of comprehensive non-clinical toxicity studies including single and repeat dose toxicity studies with twice daily dosing (BID) in rats and dogs for a duration of 28 days, in vitro and in vivo genotoxicity studies, in vitro phototoxicity studies, and in vitro and in vivo cardiovascular safety pharmacology studies. The exposure fold shown below was calculated from the predicted exposure (AUC and Cmax) at the initial clinical dose. The Cmax fold has been corrected for free fraction using a flux method using predicted species differences in protein binding and by assuming 99% protein binding for in vitro assays.

The potential of BMS-986458 to affect the cardiovascular system has been characterized both in vitro and in vivo. BMS-986458, BMT-648592, and BMT-648599 inhibited cardiac ion channels in vitro at approximately comparable concentrations, with hERG IC50 of 0.44, 0.52, and 0.16 μg/mL (0.7, 0.83, and 0.25 μM), respectively (518× for BMS-986458). BMT-648592 and BMT-648599 inhibited cardiac L-type calcium channels by 9.4% and 7.4%, respectively, at the highest test concentration of 0.628 μg/mL (1 μM). In a multi-electrode assay in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM), BMS-986458 increased field potential duration (FPDc), arrhythmia beats and decreased conduction velocity at a concentration of ≡0.19 μg/mL (0.3 μM;224×), and an increase in beat frequency was observed at 1.9 μg/mL (3 μM;2240×). In exploratory rabbit electrophysiology studies, an increase in QTc interval (friericia corrected QT [ QTcF ] = 51.2 ms;Van de Water corrected QT [ QTcV ] =24.4 ms) was noted when 12 mg/kg BMS-986458 (free plasma concentration=1.16 μg/mL,0.729 μm;1365×) was administered by Intravenous (IV) infusion, but an increase in heart rate (34%) and a decrease in mean blood pressure (-34%) interfered with the explanation. No BMS-986458 related cardiac, neurological and/or respiratory findings were noted in rats (free Cmax=0.194 μg/mL;228×) at doses up to 500 mg/kg/BID and in dogs (free Cmax=0.866 μg/mL;192×) at doses up to 50 mg/kg BID in non-GLP studies telemetry rats and dogs and in 1 month GLP rats and dogs studies.

BMS-986458, BMT-648592 and BMT-648599 were mutagenic negative in Ames assays in vitro, but positive for fragmentation and aneuploidy in the presence or absence of S9 component in exploratory in vitro assays in CHO and/or TK6 cells. BMS-986458-related effects measured on peripheral micronuclei or comets (duodenum and liver) were not noted at daily doses up to 500 mg/kg BID in non-GLP 2-week rat toxicity studies and in independent GLP-compliant in vivo genotoxicity studies, respectively, indicating that BMS-986458 is genotoxic in vivo.

Repeated dosing toxicity studies were performed in rats and dogs for a duration of up to 1 month. In key 1 month oral toxicity studies (doses of 50, 150 and 500 mg/kg BID) performed in rats, no adverse findings were present at high doses up to 500 mg/kg BID (1000 mg/kg/day total). Thus, 500 mg/kg BID was considered as a level at which no adverse effect was observed (NOAEL). BMS-986458 was orally administered at doses of 0,5, 15 and 50 mg/kg BID in a critical 1 month oral toxicity study in dogs. At a dose of 50 mg/kg BID, a decrease in food consumption was noted on day 1, leading to weight loss and a decrease in clinical symptoms, requiring early euthanasia of 3 dogs on day 7, and no clear cause of clinical state decline. All other dogs at 50 mg/kg BID survived to scheduled necropsy without significant clinical signs. No other adverse findings were present and BMS-986458 was well tolerated at doses of 15≤ 15 mg/kg BID. Thus, NOAEL and the highest non-severe toxic dose (HNSTD) were considered to be 15 mg/kg BID.

In the repeated dose toxicity study, several dose-responsive non-adverse findings were observed in rats and dogs. Immunosuppression and pro-inflammatory changes were observed in rats and dogs, including increases in circulating White Blood Cells (WBCs), neutrophils, lymphocytes, T cells, B cells, and/or Natural Killer (NK) cells, decreased production of anti-Keyhole Limpet Hemocyanin (KLH) IgM and/or IgG antibodies, decreased cytokines (interleukins [ IL ] -1β, IL-2, IL-6, tumor necrosis factor [ TNF ] α, interferon [ IFN ] γ) in rats alone, decreased amounts of lymphoid cells in thymus, intestinal-related lymphoid tissue (GALT), and lymph nodes (dogs) or spleen border areas (rats), mixed cell inflammation of femoral tibial joints in 4 dogs, and minimal neutrophil inflammation of atrial valves in 1 dog (both results were only at high doses). A decrease in Red Blood Cell (RBC) parameters with regeneration reactions was observed in both species. In rats, an increase in platelets was observed, with no consistent change in coagulation parameters in each study. An increase in bilirubin was noted in dogs, with no microscopic related abnormalities or transaminase increase, and potentially mediated by UGT1A1 inhibition. Liver hypertrophy and weight gain were observed in both species, with findings observed in rats associated with enzyme induction and secondary thyroid epithelial hypertrophy. In dogs, inconsistent changes in stool composition and positive Fecal Occult Blood (FOB) tests were observed without associated gastrointestinal pathology. Less kidney effect and no pathology-related abnormalities were noted in both species, with an increase in kidney weight in dogs and a slight increase in Blood Urea Nitrogen (BUN) in rats. Finally, minimal hypertrophy of the fascicular bands was observed in rats.

Overall, non-clinical toxicology assessment supports progression of BMS-986458 to phase 1 clinical trials.

3.2.3. Rituximab. Rituximab is a recombinant chimeric murine/human antibody directed against the CD20 antigen, a hydrophobic transmembrane protein located on normal pre-B lymphocytes and mature B lymphocytes. Following binding, rituximab triggered a cytotoxic immune response against CD20 positive cells.

Rituximab is indicated for a variety of tumor treatments and non-tumor treatments. Lymphoma indications approved in the united states and the European Union (EU) are as follows:

rituximab is indicated for use in:

treatment of patients with previously untreated diffuse large B-cell CD20 positive NHL in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) or other anthracycline-based chemotherapy regimen

Treatment of patients with previously untreated follicular CD20 positive B cell NHL in combination with CVP (cyclophosphamide, vincristine, prednisolone) chemotherapy

Treatment of patients with recurrent or refractory low-grade or follicular CD20 positive B cell NHL as single agent

European Union rituximab (MabThera) is indicated for:

treatment of patients suffering from CD 20-positive diffuse large B-cell non-Hodgkin's lymphoma in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) chemotherapy

-Treating a previously untreated patient with stage III-IV FL in combination with chemotherapy

Phase III-IV FL as monotherapy treatment chemotherapy-resistant or second or subsequent relapse after chemotherapy

-Treating adult patients with FL as a response to induction therapy as maintenance therapy

While rituximab is not approved by the FDA for the treatment of R/R lymphomas other than FL, rituximab is a component of most salvage chemotherapy regimens (e.g., R-ICE, R-DHAP, R-ESHAP, R-gemcitabine/oxaliplatin), listed in NCCN and european oncology guidelines as a treatment option for two-wire (and subsequent) treatment of B-cell lymphomas, and is routinely used in standard practice in this context. Furthermore, rituximab is approved for use in combination with lenalidomide in the united states and the european union for the treatment of patients with R/R FL.

3.3 Benefit/risk assessment

3.3.1 Risk assessment

TABLE B4-3.3.1-1 Risk assessment

Abbreviations BMA, bone marrow aspirate, BMB, bone marrow biopsy, CRBN, cerebulin, CT, computed tomography, DLBCL, diffuse large B cell lymphoma, EU, european Union, IRR, infusion-related reactions, MUGA, multiple gated acquisitions, GI, gastrointestinal tract, moA, mechanism of action, NHL, non-Hodgkin's lymphoma, PML, progressive multifocal leukoencephalopathy, PPP, prevention program pregnancy, smPC, product feature summary, TFR, oncological reactions, TLS, oncolytic syndrome, TE, thromboembolism, USPI, U.S. drug Specification, VTE, venous thromboembolism.

Standard qualification criteria are included to ensure acceptable baseline organ function (including hematopoietic function) and lack of clinically significant concomitant disease.

Since this is a FIH study, there is currently no human experience with BMS-986458. Thus, the analysis of the potential risk is based on non-clinical toxicology studies and examination of safety profiles for available compounds with similar mechanisms of action.

BMS-986458 degradation is cerebellar protein dependent, but does not lead to off-target degradation of classical CELMoD new substrates (such as Aiolos, CK 1. Alpha., ikaros, GSPT1 and SALL 4). Thus, it is expected that BMS-986458 toxicity profile will differ from CELMoD, including potential reproductive toxicity. However, since no reproductive toxicity studies have been conducted so far and the mechanism of BMS-986458 is cerebroprotein dependent, BMS-986458 will be considered a putative selective developmental poison, and participants will be required to follow BMS-986458 pregnancy prevention program (PREGNANCY PREVENTION PLAN) (PPP) to conduct BMS clinical trials.

BMS-986458 will be administered in combination with rituximab in part A2 and part B2. The most common adverse reactions (> 25%) observed in patients with NHL are infusion reactions, fever, lymphopenia, chills, infection and weakness. Fatal infusion reactions, tumor Lysis Syndrome (TLS), severe mucocutaneous reactions, and PML have been reported. The participants will also be monitored for infection and reactivation of infection (including hepatitis b reactivation), neurological function (PML), arrhythmia, and other AEs according to rituximab prescription information.

Infusion of biological products may result in severe and life threatening infusion reactions. Rituximab infusion occurred within 24 hours of the fatal infusion reaction, and about 80% of the fatal reaction occurred at the time of the first infusion (refer to USPI and EU SmPC). Including pre-medication requirements and participant monitoring.

Potential BMS-986458 and rituximab anti-cascade toxicities include cytopenia, infection, and TLS. Participants were monitored throughout the study by physical examination, AE collection, standard and specialized laboratory tests as described in this study protocol. Included in this regimen are dose adjustment guidelines, participant monitoring, and AE management guidelines.

Global 2019 coronavirus disease (COVID-19) pandemics have been identified as generally having a potential risk to clinical trial participants. Whether BMS-986458 administration would increase the risk of contracting severe acute respiratory syndrome coronavirus (SARS-CoV-2) infection or increase the severity or duration of symptoms is not known. This unknown risk must be considered when joining the participants into the group.

Because of the pandemic SARS-CoV-2, no additional safety monitoring or conventional screening tests will be required. Participants with recent or acute infections will be excluded or delayed from beginning the study intervention. If a participant suffers from a diagnosed SARS-CoV-2 infection at the time of the study intervention, it is necessary to delay administration or discontinue the study intervention.

The study was designed with study visits that allowed close monitoring of the safety of the participants throughout the clinical trial and encouraged the participants to contact the study if intermittent illness occurred between study visits. During the study, COVID-19 tests, which provide information about clinical care decisions, should follow local standard practices.

Non-live COVID-19 vaccination was considered a simple concomitant drug in the study.

3.3.2 Evaluation of benefits. Participants with R/R NHL have poor prognosis and less curative options. CA1231000 is a FIH study evaluating BMS-986458 in this patient population. Thus, while benefits may be hypothesized based on mechanisms of action and/or non-clinical observations, the actual clinical benefits of participants with advanced cancer have not been determined. The information collected during this study may help sponsors obtain information about BMS-986458 that may benefit patients with R/R NHL.

3.3.3 Overall benefit/risk conclusion. Sponsors will continue to evaluate the benefit/risk profile of the study. This assessment will be based on all available data, with particular attention to (i) AE or other safety trends in this study or any other clinical study of BMS-986458, whose characteristics, severity and/or frequency indicate that the participants will be exposed to an unreasonable and significant risk of illness or injury, and (ii) new non-clinical data indicating an unreasonable and significant risk of illness or injury.

If such an assessment indicates that the benefit/risk profile of the study is adverse to the participants, the sponsor will pause the group and/or treatment until the data is further assessed and, as to potential measures, may communicate with one or more appropriate health authorities. Such measures may include, but are not limited to, study continuation, substantial revisions, or study termination.

4. Target and endpoint

TABLE B4-4-1 targets and endpoints [ measured ]

Abbreviations AE, adverse events, AUC (0-T), area under plasma concentration-time curve, cmax, maximum concentration, CRBN, cerebulin, DLT, dose limiting toxicity, DOR, duration of response, lgG, immunoglobulin G, NHL, non-hodgkin lymphoma, ORR, total response rate, OS, total survival, PFS, progression free survival, PK, pharmacokinetics, RP2D, recommended phase 2 dose, R/R, recurrent/refractory, SAE, severe adverse events, tmax, time to maximum concentration, TTR, response time.

^a The sensitivity estimators that are mainly measured and that complement the measured will be defined in the statistical analysis plan (where applicable).

5. Study design

5.1 Overall design. This is a FIH, phase 1, multicenter, open label, dose discovery study to determine the safety, tolerability, PK, pharmacodynamics and preliminary efficacy of BMS-986458 in participants with R/R NHL. This study will be performed in part 2. Dose escalation (part a) will evaluate the escalating oral dose of BMS-986458 to evaluate safety, tolerability, and guide determination of the MTD and/or Optimal Biological Dose (OBD) of BMS-986458 as a single agent or in combination with an anti-lymphoma agent in participants with DLBCL and FL. Dose expansion (part B) the safety and preliminary efficacy of BMS-986458 administered at MTD/OBD in one or more expansion queues allocated in a randomized fashion will be further evaluated to determine RP2D.

All participants will complete screening (up to 28 days [ +3 days ]), treatment (up to 104 weeks [ about 2 years ]) (from the first dose of study intervention, with or without treatment delay) and follow-up for up to 3 study periods. The follow-up period included safety follow-up (28 days [ ±3 days after the last dose of study intervention ]), survival follow-up (up to 156 weeks [ about 3 years ] from the first dose of study intervention) and efficacy follow-up (up to 52 weeks [ about 1 year ]). The longest duration of study participation will be about 160 weeks (about 3 years) (up to 4 weeks of screening, up to 104 weeks of treatment, and up to 52 weeks of follow-up).

5.1.1. And (3) screening period. The screening period was started 28 days (+3 days of elastic window, up to 31 days) before the first dose of study intervention. Informed consent must be obtained before any study specific procedure and its completion recorded in the source file and electronic case report forms (electronic Case Report Form) (eCRF). All screening tests and procedures must be completed within 28 days (+3 days) before study treatment begins.

All participants screened will be evaluated based on inclusion criteria and exclusion criteria and will be grouped using an interactive response technology (INTERACTIVE RESPONSE TECHNOLOGY) (IRT) system. Unless otherwise stated, all screening tests and procedures must be completed within 28 days (+3 days) prior to the first dose of study intervention, and the results received and evaluated. Participants may be rescreened.

If the participant exceeds the screening period due to a study-related procedure (e.g., scheduling a tumor biopsy or waiting for a study-related laboratory value), the participant must re-agree, but a new participant identification number is not required. In this case, the minimum number of procedures for the initial screening period should be repeated to qualify the participants while maintaining the safety and qualification of the participants. Screening laboratory tests beyond the window should be repeated.

5.1.2 A treatment period. After confirmation of eligibility, the participants will enter the group and begin study intervention at the appropriate portion of the study and dose cohort.

The treatment period of BMS-986458 will last up to 104 weeks (about 2 years) from the first dose of study intervention. Details of oral administration of BMS-986458 can be found below.

For participants receiving rituximab in combination with BMS-986458, rituximab (according to the drug instructions and institutional standard practice) will be administered at a fixed dose of 375 mg/m ² for up to 6 infusions on the planned dosing day. Rituximab will be administered on day 1 of cycles 1 to 6. Details regarding rituximab infusion can be found below.

Unless otherwise indicated, any sampling or assessment should be made prior to administration on the day of visit.

5.1.2.1 Dose escalation (part a). Incremental doses of BMS-986458 will be evaluated in part A. The temporary dosing cohort for BMS-986458 dose escalation is summarized herein.

If the starting dose of BMS-986458 is intolerant, a lower dose level (i.e., cohort-1) can be explored. The temporary dose increments will follow a modified fibonacci design from 80 mg and above, where doses in successive dose cohorts can be increased by 100%, 62.5%, 53.8%, 33%, etc. of the doses in the previous cohorts. The safety review board (SRC) may recommend smaller increments of dose escalation when considering all available clinical data. SRC may open a exploratory cohort based on emerging safety, PK and pharmacodynamic data, and the starting dose will not exceed the total daily dose that has been shown to be tolerated. The SRC may propose alternative doses, schedules and frequencies. In all cases, the recommended dose for the next cohort will not increase by more than about 100% relative to the previous total daily dose.

The starting dose and schedule of BMS-986458 in part A2 will be determined by SRC based on the emerging safety, PK and pharmacodynamic data from part A1 and B1, and will be at least one dose level lower than the maximum BMS-986458 monotherapy dose explored in part B1. The initial dose of BMS-986458 in part A2 will be determined using data from a minimum of 25 participants in monotherapy treated with OBD.

Dose escalation decision. A 2-stage design will be employed in section A1 to find the MTD and/or OBD. In stage 1, BMS-986458 will be guided as a single agent dose up/down decision by Bayesian Optimization Interval (BOIN) design. The TITE-BOIN design will be used in stage 2 to find OBD. The BOIN design will be used in section A2 to guide the dose up/down decision of BMS-986458 with rituximab.

For part A1 (single dose escalation), a minimum of 3 to a maximum of 9 participants per dose level can be placed into the group cohort to assess stage 1 safety. At a target DLT rate of 30%, the dose of DLT rate within (23.6%, 35.9%) observed would be considered acceptable based on this design principle. The dose escalation/decrementing at this stage will be based on standard BOIN design methods, using the toxicity boundaries described above and setting the maximum number of participants at each dose level to 9. Phase 1 will enter phase 2 when the increment/decrement under phase 1 reaches the highest dose level within acceptable toxicity limits or the MTD is determined. At stage 2, the dosing up/down will be guided by TITE-BOIN design to assess the preliminary efficacy of each dose level and more comprehensive tolerability to determine the Optimal Biological Dose (OBD) by placing more participants into the group at the tolerable dose level determined in stage 1.

For part A2 (rituximab combination dose escalation), 2 consecutive monotherapy OBDs will be selected from part A1. Up to 16 participants (including the replenisher participants) will be treated with the selected OBD, up to 32 total participants who can evaluate the response. The combined dose escalation will be based on the same BOIN criteria as phase 1 in part A1.

The SRC will make dose escalation/decrementing, evaluate additional participants within the dose cohort, higher dose levels, use smaller dose increments, intermediate dose levels, alternate dosing schedules or frequencies, or announce final advice of MTD and/or OBD based on comprehensive assessment of safety and available PK, pharmacodynamics, and efficacy data. All recommendations presented at the SRC conference will be formally recorded (via conference summary and memo) and distributed to the researchers of the study.

The administration of one or more IMPs will be started at C1D1, after the first dose is administered in any dose level/cohort during dose escalation, the participants in each cohort are observed for a defined DLT assessment period, and then the next higher dose level can be started.

After the top 3 participants of the evaluable DLT treated in the dose escalated dose cohort completed the DLT assessment period, the tolerability of the administered dose will be determined as directed by the BOIN design and confirmed by SRC. The dose level of the next enrolled participant will be determined based on the number of DLTs observed, which describes the best dose escalation decision in stage 1 by a BOIN design with a target DLT rate of 30% (23.6%, 35.9%) and in stage 2 with the same target DLT rate and TITE-BOIN design for finding the utility matrix for OBD.

For dose levels selected for dose expansion, DLT of at least 6 participants to whom DLT can be assessed in dose increments where dose levels were determined to be tolerable. Confirmation by SRC will be based on a decision to declare tolerable dose levels based on examination of additional safety and any available PK, pharmacodynamics and efficacy data.

During the study, AE of all participants will be monitored. Adverse events will be ranked by severity according to guidelines outlined in the U.S. national cancer institute (National Cancer Institute, NCI) general term standard (Common Terminology Criteria for ADVERSE EVENTS, CTCAE) version 5.0, except for selected AEs (e.g., TLS and TFR). DLT during dose escalation will be based on AEs occurring during the DLT assessment period of C1D1 to C1D 28. Any toxicity that occurs beyond the DLT assessment period will be considered when making dose level decisions and/or dose level adjustments and defining one or more OBDs after agreement between sponsor and researcher.

Staggered administration of part a. During the up-dosing period (part a), a staggered dosing regimen will be used. The first participant in the group will be observed for at least 1 day in each dose level of more than 1 participant in the group, and then additional participants can be treated at that dose level.

5.1.2.2 Dose extension (part B). After the dose escalation is completed, the schedule of tolerable dosing may be extended in BMS-986458 OBD and/or alternatives in a plurality of queues allocated in a randomized fashion. The SRC will select the dose, schedule and treatment regimen for the expansion queue. The decision to dose extend at a given dose level and schedule will be based on, but not limited to, the overall safety profile, preliminary positive efficacy signal, and available PK/pharmacodynamic data.

Approximately 20 participants at each dose level/schedule and with sufficient follow-up will enter the B1 and B2 fractions to further evaluate the safety and efficacy of BMS-986458 alone or in combination with rituximab administered at OBD and/or lower dose levels and/or alternative schedules. Two dosage schedules will be evaluated for each indication (part B1) or each combination (part B2). The data of participants treated at OBD and/or lower dose levels and/or alternative schedules in dose escalation may be combined with the data of participants treated at the same dose levels, schedules and treatment arms in dose escalation (if from the same population). Toxicity will be monitored in monotherapy and combination expansion cohorts using bayesian continuous monitoring. The SRC will continue to monitor the overall security/tolerability data. A multi-layered approach was established to ensure adequate monitoring of participant safety throughout the study.

5.1.2.3 Stopping the definition of the standard. During part a, dose escalation will be stopped when the investigator and sponsor/medical inspector declare MTD or OBD based on the number of participants experiencing DLT events during the DLT assessment period and cumulative reviews of available safety, tolerability, PK, PD, and preliminary efficacy data.

During part B, excessive toxicity, defined as the occurrence of AEs at any time during treatment, that would meet DLT criteria specified in the dose escalation (except for dose reduction due to AEs that did not meet DLT criteria; section 10.3) will be monitored using bayesian methods. This security monitoring rule and operational features are described in section 10.2. When the stop boundary is reached, further dosing at that extended dose level will be temporarily stopped.

The mid-term calculated recommendations and all data will be reviewed by the SRC. Based on the scrutiny of the SRC, the current dose level may be permanently discontinued and lower dose levels will be evaluated in the remaining study section. Similar continuous safety monitoring will apply to the new dose level being evaluated.

In parts a and B, if any of the participants experiences any of the following Serious Adverse Events (SAE), an additional temporary SRC conference will be held for this review:

unmanageable life threatening (grade 4) events (i.e., not resolved to grade 3 or lower in the case of disruption of study intervention and medical management) unless explicitly associated with disease or external causes.

Grade 5 event (death) unless clearly associated with disease or external cause.

Any decision to review these SAE, to pause into groups or to terminate the study will be determined by the SRC.

5.1.3 A follow-up period. Follow-up will continue for all participants centering the treatment according to table B4-2-2.

5.1.3.1 Security follow-up. After the last dose of study intervention, all participants will be followed for 28 days (+ -3 days) to learn AE reports and concomitant drug information and new anti-cancer therapies, and any time thereafter SAE known to the researchers (see section 9.2.1) until recovery (return to baseline), recovery with sequelae, or death (due to SAE). The security follow-up contact may be made by telephone. One or more pregnancy tests will be required for WOCBP participants (see section 9.4.7).

All safety follow-up periods last 28 days after the last dose of study intervention (see section 9.2.5).

5.1.3.2 And (5) follow-up of the lifetime. After the last dose of study intervention, participants will be followed up by telephone contact or clinic. The survival follow-up along with the safety follow-up will be performed 28 days after the last dose of BMS-986458 and then every 12 weeks (+ -14 days) from the first dose of BMS-986458 for up to 156 weeks (about 3 years) or until death, loss of visit, withdrawal of consent or study end, whichever occurs first. In addition, information will be collected about the anticancer therapy after discontinuation of the study intervention.

The lifetime follow-up may be performed by record review (including public records) and/or telephone contact with the participant, home or treating physician of the participant.

5.1.3.3 Efficacy follow-up. Participants who discontinued study intervention for reasons other than clinically significant disease progression should continue the response assessment every 26 weeks (+ -14 days) from the last dose of BMS-986458 and up to 52 weeks (about 1 year) until disease progression, death, start of new anti-cancer therapy, loss of visit, withdrawal of consent from the study, or end of the study, subject to the first occurrence.

5.1.4.1 Security review committee. Safety considerations will be raised by the SRC, including recommendations to open a new dose level queue. The SRC will include researchers (and/or appointed representatives), sponsor's clinical trial practitioners, security practitioners, SMT chairmen, research collectiues and clinical scientists, as well as temporary participants that may include research pharmacokinetics scientists, conversion scientists and global trial managers. The SRC may consult other internal and external specialists if necessary.

The SRC will be responsible for reviewing safety data including DLT and assessing whether the dose level or schedule is tolerable or intolerable and regarding ascending/descending to subsequent dose levels, alternative doses/schedules/frequencies, intermediate doses, dose ascending using smaller dose increments, continuing, delaying or terminating dosing, repeating the queue, and entering the group of additional participants in the queue to make recommendations. The SRC will select dosages, schedules and treatment regimens for the expansion cohort and will continue to monitor the overall safety/tolerability data. All decisions made by the SRC will be formally recorded and distributed in written form to all research centers (sites). The embodiment revision will not begin dosing up, down, change the dosing schedule, or extend the existing dose cohort until a written notification of the corresponding SRC decision is sent to all participating study centers and as applicable. The final determination of the MTD and/or one or more OBDs will be determined by the SRC. One or more recommended phase 2 doses (RP 2D (s)) will be determined by the SRC based on comprehensive reviews of clinical and laboratory safety data, efficacy, and PK/pharmacodynamics/biomarkers data (as available). Before declaring one or more RP2D and consulting the researcher, the sponsor may choose to extend any dose levels previously determined to be tolerable (during DLT evaluation) in order to obtain additional experience.

5.2 Number of participants. The approximate population of participants will be 273, as follows:

dose escalation part a-approximately 73 participants can be treated, including:

o monotherapy cohort (A1 part) about 41 participants

O rituximab combination cohort (part A2) approximately 32 participants.

Dose extension part B approximately 200 participants with evaluable efficacy can be treated, including:

o one or more monotherapy cohorts (part B1) of approximately 80 participants

O one or more combined queues (part B2) of about 80 participants

One or more patient selection queues (part B3) of approximately 40 participants.

5.3 Study end definition. The start of the study was defined as the first visit of the first participant.

The primary completion day is defined as the date on which the last data point was collected for the primary endpoint of the study. If the study has multiple primary endpoints, the primary completion date is the date on which the last data point was collected for the last primary endpoint.

The end of the study is defined as the last visit of the last participant.

Participants were considered to have completed the study if they had completed all of the time periods of the study, including the last procedure shown in the activity schedule.

5.4 Study the scientific principle of design. There is a strong biological principle for evaluating BMS-986458, a new test product (IP), as a treatment for participants with R/R NHL. This phase 1, FIH clinical study of BMS-986458 will explore the safety, tolerability, PK and preliminary biological and clinical activity of BMS-986458 as a single agent or in combination with an anti-lymphoma agent in the context of recurrent or refractory disease.

5.5 Dose compliance. Dosages were selected based on the results of key 28-day toxicity studies from rats and dogs and efficacy studies in the OCI-LY1 xenograft mouse model. In a critical 28-day toxicity study in rats, all rats survived to their scheduled necropsy at doses up to the high dose, and no adverse effects were noted. In a key 28-day dog toxicity study, a related decrease in food consumption was noted in BMS-986458 at high doses. Beginning on day 1, results in clinical observations of weight loss, reduced activity, weakness, wasting, dehydration and partial eye closure, and then early euthanasia of 3 dogs on day 7. At necropsy on 3 early dead persons, there were no pathological findings indicating the cause of clinical status decline. No onset or adverse effect was noted at the medium dose, and this dose was considered to be NOAEL and the highest non-severe toxic dose (HNSTD). The following Human Equivalent Dose (HED) calculations are based on body surface area and the inter-species differential rate growth scale. STD10 in the key 28 day rat study supported the Maximum Recommended Starting Dose (MRSD) after application of the 10x safety factor. HNSTD in a key 28 day dog study supported MRSD after application of the 6x safety factor.

PK/pharmacodynamics and efficacy studies in the OCI-LY1 xenograft mouse model indicate that complete inhibition is required to achieve maximum tumor growth inhibition. Dosing was selected for evaluation in this study based on the predicted terminal half-life in humans. The initial dose in CA1231000 was chosen. Since this dose is in the middle of the predicted effective dose range and is smaller than MRSD, it is expected to be both effective and safe. Using the assumptions of predicted PK parameters and% oral bioavailability, the predicted steady state maximum plasma concentration (Cmax) and area under the concentration curve after oral administration to 70 kg humans was estimated.

The highest clinical dose will be determined based on tolerability. At the highest clinical dose presented, the minimum predicted AUC and Cmax safety margins were 15.2x and 5.6x, respectively, below NoAEL in the critical 1 month GLP toxicity study in rats or dogs.

5.6 Principle of exploratory biomarker study. In a phase 1 study evaluating BMS-986458, a series of pharmacodynamic, predictive, and exploratory biomarkers will be collected to facilitate dose/schedule selection, and potentially identify biomarkers to predict response and inform potential combinations.

More advanced image analysis techniques will be used to conduct studies relating to treatment efficacy using digital imaging samples including CT/MRI scans.

Genetic variation may affect the participant's response to the study intervention, the susceptibility to the disease, and the severity and progression of the disease. The variable response to a research intervention may be due to genetic determinants affecting the absorption, distribution, metabolism, and excretion of the intervention, the mechanism of action of the intervention, the etiology of the disease, and/or the molecular subtype of the disease being treated.

A study will be conducted using the DNA sample in association with one or more study interventions and/or diseases/disorders in the study and related disorders. Samples may also be used to develop tests/assays, including diagnostic tests related to one or more study interventions and/or diseases/conditions and related conditions in the study. Genetic studies may consist of analysis of 1 or more candidate genes or analysis of genetic markers in the whole genome or analysis of the whole genome.

All subjects participating in this study will agree with a biomarker sampling procedure, as described via ICF.

5.7 Principle of optional future research. Future studies may be conducted using residual samples initially collected from the consent participants only for another test required in the study. Future studies are intended to allow studies to be conducted on sudden or future problems that are not addressed elsewhere in the regimen, and may include studies that are not related to one or more of the study interventions and/or the disease in the study. The study may involve genetic testing using deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), which may consist of analysis of 1 or more candidate genes, analysis of genetic markers in the whole genome, or analysis of the whole genome. Analysis of samples for future studies may also involve the use of other clinical data collected as part of the study. This future study may also lead to the development of new diagnostic tests. The decision of the participant to agree to this optional future study will not affect his/her ability to participate in the primary study. Consent for optional future studies can be collected in later visits if desired. In some countries, it may require approval from local health authorities, depending on local regulations.

6. Study population. Prior approval of the protocol deviation (also referred to as protocol exemption or exemption) for the recruitment and inclusion criteria is not allowed.

6.1 And (5) selecting a standard. Participants were eligible for inclusion in the study only if all of the following criteria were applicable:

1) Signed written informed consent form

A) Participants had to sign and date written Informed Consent Forms (ICFs) approved by the regulatory, local and institutional guidelines (Institutional Review Board) (IRB)/independent ethics Committee (INDEPENDENT ETHICS Committee) (IEC). This ICF must be obtained before any regimen-related procedure is performed that is not part of normal patient care.

2) Age of participants

A) Participants must either fill 18 years of age or legal adult age (inclusive) as specified by local regulations when signing an ICF.

3) Participant type and target disease characteristics

A) Participants with a history of NHL (including DLBCL [ i.e., DLBCL NOS and diffuse large B-cell lymphoma/high grade B-cell lymphoma with DLBCL morphology with MYC and BCL2 rearrangement ] and FL) with recurrent or refractory disease according to one of the following requirements (as applicable to each indication). Furthermore, prior to group entry, the treating physician should discuss all potentially effective therapies available and available to the participants as options in terms of the best interests of the participants:

i) For R/R DLBCL (de novo) and FL 3b, after at least 2 prior normals to therapy (e.g., having received a first line combination chemotherapy regimen containing rituximab, anthracyclines, alkylating agent, and steroid, and having received at least one additional [ second line/rescue ] treatment).

Ii) for R/R DLBCL (transformed lymphoma) after at least 2 prior treatment normals (which must be administered after transformation; thus, at least 2 standard treatment regimens for DLBCL must have been accepted).

Iii) For R/R FL (except FL 3 b), the treatment criteria are met after at least 2 prior normals and upon evaluation by the investigator at the time of group entry (e.g., according to, but not limited to, the follicular lymphoma study group [ GELF ] criteria). ²¹

Iv) the participant must have a measurable disease defined by having at least one Fluorodeoxyglucose (FDG) affinity lesion for FDG affinity disease and cross-sectional imaging by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI), having a two-dimensional measurable disease and at least one lesion transverse > 1.5 cm, ²⁰ as defined by the lugano classification of NHL. Measurable disease was previously unable to be irradiated.

V) for part a, participants must agree and must have tumor biopsies and FNA-accessible during screening and FNA-accessible during cycle 1.

Vi) for part B, the participants must agree and have tumors that were accessible to paired tumor biopsies during screening and cycle 1.

4) Physical and laboratory findings

A) The following laboratory values (qualification should be confirmed prior to the first dose of study intervention) must be present:

i) Absolute neutrophil count > 1 x 109/L, and no growth factor support for 7 days (14 days in the case of pefegrelor)

Ii) an upper limit of normal value (ULN) of not more than 3.0 x or not more than 5.0 x ULN for aspartate Aminotransferase (AST) and alanine Aminotransferase (ALT) if the liver is affected

Iii) Total bilirubin is less than or equal to 1.5 x ULN

Iv) potassium, magnesium and calcium (corrected Ca or free [ ionized ] Ca) concentrations are within normal ranges or can be corrected with supplements.

V) platelet count is greater than or equal to 75 x 109/L

Vi) hemoglobin is 9 or more g/dL

Vii) estimated serum creatinine clearance of 50 mL/min or more using kidney disease diet improvement (MDRD) method

5) Reproduction state

Researchers should consult female (WOCBP) participants with fertility potential (as defined in PPP for BMS-986458) and male participants with sexual activity with WOCBP regarding the importance of pregnancy prevention, the impact of unexpected pregnancy, and the potential for fetal toxicity to occur as a result of research intervention present in semen being transmitted to the developing fetus, even if the participants have successfully performed vasectomy or companion pregnancy.

The investigator should evaluate the effectiveness of the contraceptive method in relation to the first dose of the study intervention.

Local laws and regulations may require the use of alternative and/or additional contraceptive methods.

Participants must adhere to pregnancy prevention programs.

A) Female participants:

i) Female participants without fertility potential must have evidence of record. Note that written documents may be obtained from a medical examination of a study center personnel review of the participant's medical records or medical history interviews.

(1) Women with no fertility potential (as defined in PPP for BMS-986458) are relieved of contraceptive requirements.

Ii) WOCBP must have a negative high sensitivity serum pregnancy test (lowest sensitivity 25 mIU/L or equivalent unit of human chorionic gonadotrophin) within 24 hours before BMS-986458 begins.

(1) Additional requirements of pregnancy tests before, during and after study intervention are in section 2: activity schedule.

(2) Researchers are responsible for reviewing medical history, menstrual history, and recent sexual activity to potentially reduce the risk of including women with undetectable pregnancy.

Iii) WOCBP and male participants with sexual activity with WOCBP must agree to follow the instructions of one or more contraceptive methods as described below and included in the ICF.

(1) WOCBP do not allow the use of hormonal contraceptive methods alone as a highly effective method (as described in PPP for BMS-986458).

Iv) a female is eligible for participation if she is not pregnant or lactating and meets at least 1 of the following conditions:

(1) Not WOCBP or

(2) Is WOCBP and simultaneously uses 2 reliable contraceptive regimens (a highly efficient (failure rate < 1% per year) and user independent method and an additional effective (barrier) method as described in PPP for BMS-986458) for at least 28 days before starting BMS-986458, during the intervention period and for at least 28 days after the last dose of BMS-986458, and agrees to not donate eggs (ova, oocytes) for reproductive purposes for the same period or for up to 1 year after the last dose of rituximab.

B) Male participants:

men with WOCBP sexual activity must agree to follow the instructions of one or more contraceptive methods as described below and included in the ICF.

I) Azoospermia men cannot be relieved of contraceptive requirements and will be required to always use latex or other synthetic condoms during any sexual activity with WOCBP (e.g., vaginal, anal, oral), even though the participants have undergone successful vasectomy or companion pregnancy.

Ii) male participants were asked to use the condom during the intervention period and at least 28 days after the last dose of BMS-986458.

Iii) The female partner of the male participant should be advised to use a high-efficiency contraceptive method during the study intervention period and for at least 28 days after the final dose of BMS-986458 in the male participant.

Iv) male participants with gestation or lactation partners must agree to keep prohibited activities or use the male condom during any sexual activity (e.g. vaginal, anal, oral) during the intervention period and at least 28 days after the last dose of BMS-986458.

V) during the intervention period and at least 28 days after the last dose of BMS-986458, male participants must avoid donation of sperm.

Vi) the breast-feeding partner of the male participant should be advised to consult his healthcare provider about the use of appropriate high-efficiency contraceptive measures during the time that the male participant is required to use the condom.

6.2 Criteria were excluded. Participants were excluded from the study group if any of the following criteria were applicable:

1) Medical condition

A) Participants with physical stamina equal to or greater than 3 in the eastern tumor collaboration group (ECOG) ²².

B) Participants with non-hematologic toxicity (which does not regress, return to baseline or grade 2 or are considered irreversible) associated with previous or current anti-cancer therapies.

I) After consultation sponsor medical inspectors, these participants are potentially eligible if all other inclusion/exclusion criteria are met.

C) Participants with known dysphagia, short bowel syndrome, gastroparesis, or other conditions/procedures that limit digestion or gastrointestinal absorption of orally administered drugs.

D) Participants with uncontrolled hypertension (systolic BP > 180 mm Hg or diastolic BP > 100 mm Hg) or hypotension (systolic BP < 90 mm Hg or diastolic BP < 60 mm Hg) at screening or within 1 month before C1D 1.

E) Clinically significant cardiovascular or cerebrovascular disease, including any of the following:

i) Complete left or double bundle branch block or other clinically significant abnormal ECG findings at screening.

Ii) history of congenital long QT syndrome.

Iii) Sustained or clinically significant ventricular arrhythmias were screened.

Iv) the Fridericia formula was used to screen ECG for a QT interval (QTcF) of 470 milliseconds for heart rate correction.

V) a history of unstable angina, myocardial infarction or stroke or clinically significant cardiac arrhythmias (such as ventricular tachycardia, ventricular fibrillation or torsades de pointes) less than or equal to 6 months prior to the initiation of the study intervention.

Vi) the cardiovascular function impaired state of the New York Heart Association is greater than or equal to grade 3 ²³ during screening. Grade 3 is defined as heart disease that results in significantly limited physical activity and participants are comfortable at rest, but less than normal physical activity can lead to fatigue, palpitations, dyspnea, or angina.

Vii) LVEF < 45% at screening, as determined by Echocardiography (ECHO) or multi-gated acquisition (MUGA) scans.

F) Participants with any disorder that interferes with the ability to interpret data from the study.

G) Participants in any condition that would place the participants at unacceptable risk if participating in the study, including significant acute or chronic medical conditions, active or uncontrolled infections, or the presence of laboratory abnormalities.

H) Participants with known SARS-CoV-2 infection with mild or asymptomatic infection within 10 days or severe/critical illness within 20 days prior to cycle 1, day 1. The acute symptoms must have resolved and, based on the assessment of the researcher, there is no sequelae that put the participant at high risk of receiving research intervention after consulting the sponsor medical inspector.

I) Venipuncture is not possible and/or venous access is not tolerated.

J) As determined by the researcher for any other reasonable medical, mental and/or social reason.

K) A concurrent history of the second cancer requiring ongoing sustained systemic treatment.

L) participants with a history of known or suspected Central Nervous System (CNS) involvement or primary CNS lymphomas (including those in remission). Only participants with suspected brain involvement during screening need to undergo brain/spinal scans (CT or MRI).

2) Reproduction state

A) Pregnant or lactating females.

3) Previous/concomitant therapy

A) Treatment such as the limitations, precautions, and banned listed in section 7.7 concomitant therapy cannot be followed.

B) Live virus vaccine was previously live for less than or equal to 4 weeks before starting the intervention.

C) The previous COVID-19 vaccine was within 7 days prior to the beginning of the intervention. For vaccines requiring more than 1 dose, when applicable and when the delay in initiating a study intervention does not put the study participants at risk, then the full series (e.g., two doses of a2 dose series) should be completed at least 7 days prior to initiating the study intervention.

D) Participants had received prior systemic anti-cancer therapy (approved or experimental) within less than 5 half-lives or 4 weeks (whichever was shorter) before beginning the study intervention

E) Participants had received prior CAR-T or other T cell targeted therapies (approved or experimental) within +.4 weeks prior to initiating study intervention.

F) The participants had received prior therapy with CRBN modulating drugs (e.g., lenalidomide, atorvastatin, pomalidomide, CC-99282) within +.4 weeks before starting the study intervention.

G) Participants are receiving long-term systemic immunosuppressive therapy or corticosteroids (e.g., prednisone or equivalent over 10 mg/day over the last 14 days) or participants with clinically significant Graft Versus Host Disease (GVHD).

I) Allowing stable use of inhaled corticosteroids.

Ii) for ongoing skin or eye GVHD, topical steroids are allowed.

H) Participants received prior autologous SCT within less than or equal to 3 months prior to beginning study intervention. If the participants received prior autologous SCT within > 3 months prior to the beginning of the study intervention, then any treatment-related toxicity did not resolve (> grade 2).

I) Participants before beginning study intervention

And less than or equal to 6 months of prior allogeneic SCT that has been subjected to standard or reduced strength pretreatment. If the participants are before beginning the study intervention

> 6 Months of prior allogeneic SCT received, > no treatment-related toxicity resolved (> grade 2).

J) Participants received major surgery less than or equal to 2 weeks before the study intervention began. The participants must have recovered from any clinically significant effects of recent surgery.

K) Prior radiation therapy was not more than 4 weeks prior to the beginning of study intervention.

4) Physical and laboratory test findings

A) At the discretion of the investigator, there is evidence that there is organ dysfunction or any clinically significant deviation from normal (which is outside of the range consistent with the target population) in physical examination, vital signs, ECG, or clinical laboratory determinations.

B) Participants with known HIV seropositive or active infection, or active infection with Hepatitis B Virus (HBV) or hepatitis C virus (as evidenced by the positive screening of blood for hepatitis C antibodies; hepatitis B surface antigen and anti-hepatitis B surface antibodies, or HIV-1 and HIV-2 antibodies).

5) Allergy and adverse drug reactions

A) History of allergy to rituximab or any excipient.

6) Other exclusion criteria

A) Prisoners or involuntarily banked participants. ( Note that in certain specific situations and only in countries where local regulations allow, caged persons may be included or allowed to continue as participants. Stringent conditions apply and require sponsor approval. )

B) Is involved in another clinical trial conducted concurrently with this study.

C) Blood pools were donated 4 weeks prior to the study intervention (for plasma only, within 2 weeks) or in clinical studies (except at screening visit).

6.4 Screening failed. Screening failure is defined as the participants who agree to participate in the clinical study but subsequently did not enter the group study/inclusion analysis population.

A minimal set of screening failure information is required to ensure transparent reporting of screening failure participants, meet test report unification criteria (Consolidated Standards of Reporting Trials, CONSORT) issue requirements (as applicable), and respond to the queries of regulatory authorities. Very little information includes date of consent, demographics, details of screening failure, qualification criteria, and any Serious Adverse Events (SAE) that occur after consent.

6.4.1 Retests during screening. This study allowed for re-entry of participants who discontinued the study due to screening failure (i.e., participants had not been randomly assigned/received treatment). If the group is re-entered, the participants must re-agree.

It will allow laboratory parameters and/or other evaluations to be retested in any single screening (in addition to any parameters that require validation values).

The latest result before the first dose of study intervention is the value by which the assessment study was incorporated, as it represents the latest clinical state of the participants.

The laboratory parameters and/or evaluations included in the screening program summary (SCREENING PROCEDURAL OUTLINE) may be repeated in an effort to find all possible fully qualified participants. It may be necessary to consult a medical inspector to identify whether repeated testing of any particular parameter is clinically relevant.

7. One or more research interventions and concomitant therapies. Study intervention is defined as any one or more trial interventions, one or more marketed products, placebo, one or more procedures, or medical devices intended to be administered to study participants according to a study regimen.

Research interventions included trial [ drug ] products (IP/IMP) and non-trial/co [ drug ] products (non-IP/non-IMP/AxMP).

Test product (IP), also referred to as test drug (IMP) in some areas, is defined as a pharmaceutical form of active substance or placebo tested or used as reference in clinical studies, including products that have been marketed but are used or assembled (formulated or packaged) differently from the authorized form, products for unauthorized indications, or products when used to obtain further information about the authorized form.

Other drugs used as support drugs or salvage drugs (escape medication) for prophylactic, diagnostic, or therapeutic reasons as components of standard care for a given diagnosis may be considered non-IMP/AxMP.

7.1 Study intervention of administration. The duration of the treatment period was 28 days. Study visits will be performed as detailed in the table of activity schedules. BMS-986458 will be administered according to a prescribed schedule for up to 104 weeks (about 2 years) from C1D1 (regardless of dosing delay) or until progression, unacceptable toxicity, or withdrawal is decided by the participant or physician. Rituximab will be administered IV on day 1 from cycle 1 to cycle 6.

See information about dose adjustment, treatment discontinuation and safety management, respectively.

TABLE B4-7.1-1 study intervention

Abbreviations IMP, test drugs, smPC, product characteristics summary.

^a If allowed by local regulations, these products are available in some countries as local commercial products from research centers. In these cases, the package size/efficacy of the product may be different from that listed in the table. These products should be prepared/stored/administered according to the pharmaceutical instructions or SmPC.

BMS-986458 was administered. BMS-986458 will be administered orally continuously from cycle 1, day 1 (C1D 1). The schema is revised before any other alternative schedule is introduced.

BMS-986458 will be administered at the clinic on the study day where PK samples will be collected.

Participants will be required to record the date and time of each self-administration study intervention. Participants in all groups were asked to fasted (not to eat or drink anything other than water) about 2 hours before administration of BMS-968458 up to 1 hour after. In some cases, participants may be allowed to avoid food and beverage (other than water) restrictions 2 hours before and 1 hour after administration of BMS-968458 after discussion with a medical inspector.

If the participants vomit within 30 minutes of the BMS-986458 intake, the BMS-986458 tablet may be taken again. If emesis occurs after 30 minutes of BMS-986458, the participants must not ingest additional BMS-986458 tablets unless the complete medication is visible in the vomit.

BMS-986458 can be administered for up to 104 weeks (about 2 years) or until clinically significant disease progression, unacceptable toxicity, or withdrawal of consent.

At the discretion of the researcher and after discussion with the medical inspector, retreatment with BMS-968458 may be considered if all of the following criteria are met:

Participants discontinue treatment for reasons other than AE, progression, or initiation of new anti-lymphoma therapy

Based on the malignant lymphoma International working group criteria (The International Workshop Criteria for Malignant Lymphoma), participants achieved a demonstrated response with BMS-986458

Based on the international working group criteria for malignant lymphoma, participants have evidence of disease recurrence during the lifetime follow-up period

Continue to qualify

Participants receiving retreatment will follow the schedule of activity starting on day 1 of cycle 1, but may continue with their most recent dose level and a schedule or dose level at which at least one participant cohort in the study shows adequate tolerance. The participants were treated only once more during the study.

For further details of storage, preparation and administration of BMS-986458, please refer to the current manual of researchers and/or pharmaceutical manuals.

7.1.2 Rituximab is administered. Rituximab will be administered at a dose of 375 mg/m ² at day 1 of cycles 1 to 6 as an IV infusion. Dose reduction of rituximab is not allowed.

Rituximab may be administered with a morning dose of BMS-986458 or within up to 2 hours after a morning dose of BMS-986458. Suggested body surface area estimation methods include Mosteller ²⁴ and DuBois ²⁵.

Bsa= v ((height (cm) x weight (kg))/3600

Bsa= 0.20247 x height (m) 0.725 x weight (kg) 0.425

Preparation, infusion rates and pregnancy prevention guidelines should be made according to locally approved rituximab prescription information or local institutional practices (rituximab PI). BSA calculation methods and time settings will be performed according to institutional guidelines, and prior to each rituximab infusion, a precursor dosing consisting of acetaminophen and an antihistamine should be administered (see current local rituximab prescription information). Depending on institutional practices, steroids may also be administered prior to the onset of rituximab infusion. Monitoring measures during and after infusion of rituximab should be applied as recommended by the manufacturer and current guidelines.

Participants who do not require dose interruption are allowed to enter the next treatment cycle (e.g., hematologic toxicity must resolve to < 2). If rituximab doses are discontinued due to study intervention-related toxicity, rituximab may be restarted once toxicity subsides to < grade 2. In cycle 1, the skipped dose due to toxicity will not be rescheduled. In a period of ≡2, the missing dose of rituximab can be made up within 2 weeks (i.e. days 1 to 14 of the period). If toxicity subsides to <2 orders during days 15 to 28 of the cycle, rituximab dosing will resume on day 1 of the next cycle.

Participants will be closely monitored throughout the infusion period and for at least 1 hour after continuous infusion, and a series of vital signs are obtained and pulse oximetry monitoring is performed. Complete emergency resuscitation facilities should be immediately available and participants should always be under close supervision of a researcher or appropriately trained staff.

In participants with grade 4 infusion reactions or recurrent grade 3 infusion reactions requiring permanent rituximab discontinuation, additional blood tests can be obtained within 15 to 60 minutes of the onset of symptoms to assess severe allergic reactions. Laboratory tests may include immunoglobulin E (IgE), total tryptase, and plasma histamine.

The medical inspector should be informed of any grade 4 or recurrent grade 3 infusion reactions within 24 hours of the event.

For additional details of dosing and stored information, reference is made to locally approved rituximab tags or pharmacy manuals.

7.2 Assigned to study intervention. All participants will use IRTs centrally. Before the study starts, each user will receive login information and directions on how to access the IRT. If desired, the research center will record treatment assignments on an applicable electronic case report table (eCRF). During the screening visit, the study center will assign a 5 digit number of participants using the IRT, which is unique at all study centers. Participants in the group (including those not dosed) will be assigned consecutive participant numbers. Group entry participants meeting inclusion criteria and exclusion criteria will be eligible to receive dosing. The sequential numbering of each participating study center may be restarted because the different patient identification numbers will ultimately include the study center number and the participant number. After all screening procedures are completed, the research center will either screen for failure using IRTs or obtain treatment assignment information, including random assignment (as applicable) to an extended cohort. Subsequent visits will be registered in the IRT system for drug delivery. If the IRT system is not available, the participant will be manually entered into the group.

Study interventions will be assigned at study visits listed in the activity schedule.

7.3 And (5) blind people are arranged. This is an open label study (open to sponsors and study centers/participants; not treated with sponsor blindness). It has been determined that blinding is not required to meet the study objectives. Blinding procedures are not applicable and access to treatment allocation information by any Bristol-Myers quick company (BMS) team, study center or participants is not limited. IRTs will be used to assign the particular treatment that the participant receives. The research center will contact the interactive response system (INTERACTIVE RESPONSE SYSTEM) prior to the initiation of the study intervention administration for each participant. If desired, the research center will record treatment assignments on an applicable electronic case report table (eCRF).

7.4 Dose adjustment

7.4.1 Dose adjustment. The guidelines for dose adjustment of BMS-986458 are presented in tables 7.4.1-1 and 7.4.1-2 for study intervention-related AE. These cases should be discussed with a medical inspector before a dosing change is made. Only 2 dose reductions are allowed before a discontinuation is required, or only 1 dose reduction if at the starting dose. If BMS-986458 is a permanent study intervention-related AE. These cases should be discussed with a medical inspector before a dosing change is made. Only 2 dose reductions are allowed before a discontinuation is required, or only 1 dose reduction if at the starting dose. If BMS-986458 is permanently suspended in any combination queue, the combination anti-lymphoma agent should also be suspended.

For rituximab-related toxicity, recommendations follow the drug instructions and local practices. Dose reduction of rituximab is not allowed.

For AEs not summarized in tables 7.4.1-1 and 7.4.1-2, they are discussed with medical inspectors.

TABLE B4-7.4.1-1 guidelines for the dosage adjustment of hematological toxicity

Abbreviations ANC, absolute neutrophil count, CBC, whole blood count, DLT, dose limiting toxicity, G-CSF, granulocyte colony stimulating factor, GM-CSF, granulocyte-macrophage stimulating factor.

TABLE B4-7.4.1-2 guidelines for dose adjustment of non-hematological toxicities

Abbreviations AE, adverse events, ALT, alanine aminotransferase, AST, aspartate aminotransferase, ANC, absolute neutrophil count, DLT, dose limiting toxicity, ECG, electrocardiogram, IV, intravenous, NCI CTCAE, american national cancer institute adverse events generic term standard (National Cancer Institute Common Terminology Criteria for ADVERSE EVENTS), NSAIDs, non-steroidal anti-inflammatory drugs, QTc, corrected QT, QTcF, qtinterval corrected by FREDERICIA.

7.4.2 Management of selected adverse events

7.4.2.1 Infection. Vigilance and management of signs and symptoms of infection should be practiced according to institutional standard medical practice. Conventional infectious disease prevention is not recommended, however, antibiotics, antiviral, pneumosporoal resistance, antifungal or other preventive measures may be administered at the discretion of the researcher and/or SRC during the study. For guidelines for dose adjustment, refer to tables 7.4.1-2.

7.4.2.2 SARS-CoV-2 infection. Participants diagnosed with SARS-CoV-2 infection should discontinue one or more study interventions and the study interventions can be resumed when no active disease signs are present and confirmed by a researcher evaluation. Allowing for treatment of active SARS-CoV-2 infection or high risk exposure, including using experimental therapies and taking into account the risk of DDI (e.g., pampers Luo Weide (Paxlovid)), and should be discussed with medical inspectors.

7.4.2.3 Management of Tumor Lysis Syndrome (TLS). Participants considered at risk for TLS (e.g., having a high tumor burden or high circulating lymphocyte count [ > 25 x 10 ⁹/L ]) must begin to receive adequate tumor lysis prevention and hydration with an anti-uric acid agent (e.g., allopurinol) 12 to 24 hours before BMS-986458 begins. For treatment of TLS, electrolyte abnormalities are corrected, renal function and fluid balance are monitored, and supportive care, including dialysis, is administered, as indicated. Researchers will make clinical decisions as to whether more frequent serum chemistry monitoring is appropriate.

For the TLS definition, reference is made to the Tumor Lysis Syndrome (TLS) prevention recommendation ²⁶, the Cairo-Bishop definition of laboratory TLS, and the Cairo-Bishop classification system ²⁷ of TLS.

7.4.2.4 A tumor burning reaction (TFR). The participants in this study should be monitored for neoplasia. A oncological reaction (TFR) is defined as a sudden and painful increase in size of the affected area including lymph nodes, spleen and/or liver, often accompanied by low grade fever, non-pruritic diffuse rashes and in some cases an increase in peripheral blood lymphocyte count. The burning tumor generally occurs during the first cycle of treatment. The oncological response may mimic the progression of the disease, however, TFR generally resolves over time. Thus, careful monitoring and evaluation is important before the participants are discontinued due to disease progression in the initial cycle of BMS-986458.

There are currently no laboratory or radiological tests that distinguish TFR from disease progression. Differentiation should be made before and after initiation of treatment based on clinical basis of combined observations such as relevant body findings, laboratory findings and disease progression rate. The burning tumors will be recorded as AE and graded using NCI CTCAE 3.0 standard.

Treatment of TFR was judged by researchers based on severity and clinical status. In mild to moderate cases (grade 1 to 2), it is recommended to continue BMS-986458 with symptomatic treatment, such as NSAIDs and/or anesthetics. The administration of the corticosteroid may be at the discretion of the researcher. In more severe cases (grade 3 to 4), BMS-986458 should be discontinued and corticosteroid therapy should begin. Researchers may also decide to use NSAIDs and/or anesthetics. When symptoms subside to < 1 level, BMS-986458 can be restarted at the same dosage level. For examples of possible TFR, researchers are encouraged to discuss with sponsor medical inspectors.

7.4.2.6 Infusion-related reactions. Administration of rituximab may result in severe (including fatal) infusion-related reactions. A fatal infusion reaction occurred within 24 hours of rituximab infusion, with about 80% of the fatal infusion reaction occurring in association with the first infusion. Symptoms may include fever, chills/tremors, nausea, itching, angioedema, hypotension, headache, bronchospasm, urticaria, rash, vomiting, myalgia, dizziness or hypertension. Infusion-related reactions typically occur within 30 to 120 minutes of the start of the first infusion of rituximab and subside by slowing or interrupting infusion and supportive care (diphenhydramine, acetaminophen, and intravenous saline) (see US PI and/or SmPC).

Including pre-medication requirements and subject monitoring. If a severe and life threatening infusion reaction occurs, complete emergency resuscitation facilities should be immediately available and the subject should be under close supervision of the researcher or appropriately trained staff during the infusion. Medical management of infusion reactions should be enabled according to standard medical practice using glucocorticoids, epinephrine, oxygen, intravenous infusion, and other drugs (according to medical indications).

7.4.2.7 Progressive multi-range leukoencephalopathy. John Cunningham (JC) virus infection leading to Progressive Multifocal Leukoencephalopathy (PML) was observed in participants treated with rituximab, which can be fatal. The PML of a participant with a new nervous system manifestation or a preexisting nervous system manifestation with a change should be evaluated.

Symptoms of PML are non-specific and may vary depending on the affected area of the brain. Motor symptoms are common with findings of corticospinal tracts (e.g., muscle weakness, paralysis and sensory disturbance), paresthesias, cerebellar symptoms and visual field defects. Some signs/symptoms may occur that are considered "cortical" (e.g., aphasia or visual space disorientation). Evaluation of PML includes, but is not limited to, consultation neurologists, brain MRI, and lumbar puncture (cerebrospinal fluid test for john cunningan virus DNA). Rituximab and BMS-986458 should be discontinued in the participants with PML. The participants should be forwarded to the neurologist for evaluation and treatment of PML.

7.4.3 BMS-986458 and rituximab dose interruption criteria. Unless otherwise stated in tables 7.4.1-1 and 7.4.1-2, participants experiencing the following should discontinue study intervention.

Any study intervention related AE that meets the definition of DLT will require discontinuation of dosing. If AE did not improve to < 2 orders or baseline, this may require a subsequent dose reduction. Such cases should be discussed with sponsor medical inspectors to determine the optimal duration of administration delay or reduction.

Investigation of intervention-related ≡3 grade toxicity, chronic grade 2 toxicity or significantly missed doses due to tolerability may require dose reduction of BMS-986458. Such cases should be discussed with a medical inspector before a drug administration change is made.

Confirm SARS-CoV-2 infection (see section 7.4.2.2).

Treatment with one or more study interventions can be discontinued for up to 4 weeks until toxicity reaches < grade 2 or baseline levels. In rare cases, participants may benefit from recovering BMS-986458 after an interruption longer than 4 weeks. All decisions about whether to resume administration after > 4 weeks of disruption must be discussed with the medical inspector. BMS-986458 can be restarted with the same dose or a reduced dose as described in section 7.4.3.1.

In part a during cycle 1, a disruption of study intervention for reasons other than DLT for > 7 days would render the DLT of the participant non-evaluable and require replacement of the participant in the dosing queue. Any such interruption must be discussed with the sponsor medical inspector.

Even if dosing is delayed, evaluation continues on schedule.

7.4.3.1 Restoring the standard of treatment. Unless otherwise stated in tables 7.4.1-1 and 7.4.1-2, once non-DLT AEs associated with study intervention subsided to ∈1 or baseline, participants experiencing AEs that did not meet permanent discontinuation criteria may resume treatment with one or more study interventions. Medical inspectors/designated personnel must be consulted before study intervention is resumed in participants who have had their dosing discontinued for > 4 weeks.

7.4.3.2 Dose reduction criteria. The dose of BMS-986458 is allowed to decrease during any period (including period 1) after the consultation sponsor medical inspector. The BMS-986458 dose reduction due to AEs occurring in cycle 1 during dose escalation will constitute DLT (see section 7.4.1), but will allow the participants to continue to receive BMS-986458 at the reduced dose.

When a dose reduction of BMS-986458 is indicated, the next lower dose level deemed safe and tolerable will be selected. If the participant is on a dosage schedule, the participant can reduce the dosage to the total daily dose equivalent of the next lower dosage level administered in the schedule. For part B, this means that the participant can reduce the dose to the total daily dose equivalent of the next lower dose level in part a that is considered safe and tolerable, which is administered on the same schedule as the participant is dispensed. Once the dose is reduced, the dose may be increased when the toxicity reaches < 1 grade. If toxicity recurs at higher doses, the dose will be reduced a second time, but then not allowed to increment again. Two dose reductions were allowed (one for the starting dose). If any of the participants continued to experience unacceptable toxicity after 2 dose reductions (one reduction was for the starting dose), BMS-986458 would be permanently discontinued.

Chronic grade 2 toxicity may require dose reduction of the study intervention. Such cases should be discussed with a medical inspector before making the change.

Dose reduction is not allowed for rituximab.

7.4.3.3 Dose escalation criteria. During cycle 1, the intra-participant dose is not allowed to escalate beyond the dose initially allocated to the participant. Participants in part a who were decided by the investigator to continue taking BMS-986458 from cycle 2 after SRC approval may have increased dose levels provided that alternative dose levels have been shown to be well tolerated by other participants in at least 1 cohort in this study. To account for intra-participant dose escalation, participants must exhibit adequate tolerance to their current dose level (e.g., not experience BMS-986458-related ≡3-grade toxicity at their prescribed dose level). In part B, dose escalation beyond MTD is not allowed.

7.4.4 Definition of dose limiting toxicity. Toxicity severity will be graded according to NCI CTCAE version 5.0, except TLS and TFR, which will be graded according to Cairo-Bishop TLS grading system ²⁶ and NCI CTCAE version 3.0, respectively.

The DLT evaluation period is defined as cycle 1, day 1 to cycle 1, day 28, including pre-dose allocation prescribed for cycle 2, day 1. Any event will be considered a DLT unless the researcher can attribute the event to a clearly identifiable cause, such as underlying illness or disease progression, other concurrent illness or concomitant medication.

Dose limiting toxicities are described below:

Hematological toxicity is as follows:

o exothermic property of any duration neutropenia

Grade 4 neutropenia lasting > 7 days

O grade 4 thrombocytopenia or grade 3 thrombocytopenia with > 24 hours of duration with clinically significant bleeding

O 4-stage anemia lasting > 24 hours

Any non-hematologic toxicity of grade 3 or more, except for the following:

grade o3 nausea, vomiting, dehydration or diarrhea, which subsides within 72 hours (with optimal medical management) and does not require feeding tubes, total parenteral nutrition or prolonged hospitalization.

O grade 3 IRR controlled by study intervention discontinuation and medical management, which subsided to grade 1 or less within 48 hours.

Grade o3 fatigue, which subsided to grade 2 less than 7 days of discontinuation of study intervention and did not relapse at the same level after recovery of study intervention at the same dose (with optimal medical management).

Grade o3 TLS if it has no clinical complications and regresses with routine medical management within 7 days, and no end organ damage.

Grade o3 electrolyte abnormality (i.e., electrolyte abnormality that occurs without clinical consequences) subsides to a grade 2 level within 72 hours with or without intervention.

O3 grade AST or ALT increases, which subsides to < 2 grade within 72 hours and does not meet the standards of the Hai's law (Hy ́ s law).

Grade o3 acneiform, pustular or maculopapular rash resolved to grade 2 (with best medical management) within 7 days of discontinuation of study intervention. For symptom control, topical steroid treatment is allowed.

Any confirmed cases of Hai's law. Note that Hai's rule cases are cases where liver transaminase levels ≡3×ULN is accompanied by an increase in total bilirubin to > 2×ULN, but without initial findings of cholestasis (e.g., elevated serum ALP), no other reason could be found to explain the increased combination of transaminase and total bilirubin.

Any AE, unless specifically determined to be not relevant to one or more study interventions, and a dose level reduction during cycle 1 is required.

Any death during cycle 1.

Isolated laboratory changes that are devoid of relevant clinical signs or symptoms and/or ECG changes (e.g., hypomagnesemia, hypermagnesemia, hypoalbuminemia, hypophosphatemia, increased or decreased lymphocyte count) may not be included in this definition. These findings will be discussed and reviewed by the SRC.

Any toxicity that occurs outside of the DLT period will be considered in making the final dose level determination.

For measures taken against BMS-986458 and rituximab with respect to DLT or other AEs, see the dose adjustment guidelines (see tables 7.4.1-1 and 7.4.1-2).

7.4.5 The definition of participants in DLT can be evaluated. All participants receiving at least 1 dose of BMS-986458 will be rated for safety. After the first dose is administered in any of the participant cohorts during dose escalation, the participants in each cohort will be observed for 28 days (DLT assessment period) and then the next higher, regimen-specified dose schedule cohort may be started.

Participants that can evaluate DLT are defined as those who receive at least 75% of the one or more IMPs of the total planned cycle 1 dose intensity and do not experience DLT during the DLT evaluation period.

Or (b)

Participants who experienced DLT during the DLT assessment period after receiving at least one dose or a part of one or more IMPs thereof.

Participants that cannot evaluate DLT may be replaced. Additional participants in any tolerating dose cohort may be decided by the researcher and enrolled after an agreement with the sponsor medical inspector.

7.4.6 Determination of MTD, one or more OBDs, and one or more RP2 ds. MTD is defined as the dose level that gives a probability that can bring the estimated DLT to the nearest about 30%. The preliminary estimate of MTD will be based on an estimate of DLT incidence at each dose for the participants of evaluable safety of all treatments in phase 1 and phase 2, and OBD will be estimated based on the TITE-BOIN (phase 2) algorithm. The final determination of MTD and OBD will be decided by SRC.

Based on cumulative safety, PK/pharmacodynamics (as available) and efficacy data from participants in the treatment in part a, 1 or more dose levels can be selected for further evaluation in part B. One or more RP2D will be recommended by the SRC after examining the data from parts a and B of the study.

7.5 Preparation/processing/storage/check registration (accountability). According to local regulations, IP/IMP/non-IMP/AxMP must be stored in a secure area. The responsibility of the researcher or the allowed designated personnel is to ensure that IP/IMP/non-IMP/AxMP is assigned only to research participants. According to local regulations, IP/IMP/non-IMP/AxMP must be distributed from the official study centre only by authorised personnel.

The product storage manager should ensure that research interventions are stored according to the environmental conditions (temperature, light and humidity) as determined by the sponsor. If problems arise with respect to the quality or appearance of the research intervention, the research intervention should not be distributed and the sponsor should be contacted immediately.

Research interventions not supplied by sponsors will be stored according to the drug instructions.

The IP/IMP/non-IMP/AxMP written document (whether supplied by the sponsor or not) must be maintained and must include all the procedures required to ensure accurate administration of the drug. This includes written documents of drug storage, administration, and (if applicable) storage temperature, reconstitution, and use of the desired method (e.g., diluent, applicator as desired).

Researchers or prescribing personnel must confirm that all study interventions accepted during transportation maintain appropriate temperature conditions and report and resolve any discrepancies prior to use of the study interventions.

Only participants in the group study may receive the study intervention, and only authorized study center staff may supply, prepare or administer the study intervention.

Depending on the storage conditions of the markers, all research interventions must be stored in a safe, environmentally controlled and monitored (manual or automatic) area, which is limited to contact by researchers and authorized research center staff.

Researchers, institutions, medical institution authorities (as applicable) or authorized research center personnel are responsible for research intervention check registration, reconciliation and record maintenance (i.e., receipt of records, reconciliation and final disposition).

Further guidance and information on the final treatment of unused study interventions is provided in the pharmaceutical handbook.

For research interventions not provided by sponsors but commercially available from research centers, the storage should be based on product labels.

7.6 Intervention compliance was studied. Participants will be required to record their daily BMS-986458 self-administration at home in the dosing diary. Study intervention compliance will be monitored periodically by medication check-in (audit administration diary). Drug accountability should be reviewed by study center staff at each visit to confirm treatment compliance. Study center staff should discuss differences with participants at study visits in each treatment. Compliance will be assessed by directly asking and counting the back tablets during the study center visit and recorded in the source file and related tables. One or more deviations from the prescribed dosing regimen should be noted in the eCRF.

The start and stop dates of study intervention, including the date of intervention delay and/or dose reduction, will be recorded in eCRF. A record of the number of BMS-986458 tablets allocated to and taken by each participant must be maintained and checked against the study intervention and compliance records.

7.7 Concomitant therapy

7.7.1 Prohibited and/or limited therapy. Prohibited and/or limited drugs taken prior to administration of the study intervention are as follows:

prior exposure to BMS-986458

Exposure to any trial drug or placebo during the study period within 4 weeks of study intervention

Anticancer therapies (chemotherapy, biological or experimental therapies, and surgery) other than study intervention during the study. After discussion with sponsor medical inspectors, a researcher decides that focal palliative radiation therapy may be allowed for treatment of symptoms associated with cancer treatment during the course of research treatment. Medical inspectors will have the opportunity to provide advice to research center researchers regarding drug elution prior to radiation therapy and allow whether to resume treatment based on possible radiation toxicity. Maintenance of sex hormone therapy is permitted in subjects with a history of breast or prostate cancer.

Supplementary drugs (e.g., herbal supplements or traditional Chinese medicines) should be carefully used. The use of san John's grass (St. John's woli) is not allowed, see the list of prohibited CYP3A inducers and inhibitors below.

Any live/attenuated vaccine (e.g., varicella vaccine, shingles vaccine, yellow fever vaccine, rotavirus vaccine, oral polio vaccine, and measles, mumps, rubella [ MMR ] vaccine) during the study and up to 28 days after the last dose of study intervention.

O allows for an attenuated vaccine or an inactivated vaccine or a component vaccine, such as an injectable annual influenza vaccine.

O no live COVID-19 vaccine should be administered to the participants during the study (including during the treatment) and during the 28 day follow-up period following the last dose of study intervention. In addition, administration of live COVID-19 vaccine was prohibited up to 28 days prior to initiation of study intervention.

Other experimental therapies during the study are used, except therapies for treating active SARS-CoV-2 infection or high risk exposure that are allowed and should be discussed with medical inspectors. The administration of the SARS-CoV-2 vaccine for testing is not allowed.

Strong CYP3A4 inhibitors and inducers, sensitive CYP3A substrates or CYP3A substrates with a narrow therapeutic window.

Examples of strong CYP3A inhibitors, inducers, and substrates include (but are not limited to):

o atazanavir, sevilia orange

O-alfentanil, alvardill, buspirone, colpitan, darifenacin, darunavir (darnnavir), ebastine, everolimus, ibrutinib (ibrntinib), lometapi (lornitapide), lovastatin, midazolam, naloxonol (naloxegol), nisoldipine, saquinavir, simvastatin, sirolimus, tacrolimus, telanavir, triazolam, vardenafil, budesonide, dasatinib, dronedarone, eletriptan (eletriptan), eplerenone, felodipine, indinavir, lurasidone, malavirol, quetiapine, sildenafil, ticagrelor tolvaptan

In vitro solubility data indicate that the solubility of BMS-986458 decreases with increasing pH. Concomitant use of proton pump inhibitors and H2 antagonists was prohibited until further data was available. If it becomes medically necessary at the time of the study (e.g., peptic ulcer disease, gastroesophageal reflux disease, erosive esophagitis), they can be carefully given after discussion with a medical inspector. Short-acting antacids such as Maalox or TUMS may be administered, but it is recommended that these are not administered 2 hours prior to or 2 hours after BMS-986458 administration.

Participants should consume seville orange, grapefruit or grapefruit juice, grapefruit, exotic citrus fruits, grapefruit hybrids or fruit juice from 5 days prior to the start of study intervention until 24 hours after the final dose of BMS-968458.

Drugs taken within 4 weeks prior to study intervention and up to 28 days after the last dose of study intervention must be recorded on eCRF.

7.7.2 Allowed therapies. The participants were allowed to use the following treatments when receiving study intervention:

At the discretion of the researcher, granulocyte colony-stimulating factor (G-CSF) or granulocyte-macrophage colony-stimulating factor (GM-CSF) for therapeutic purposes

Parenteral influenza vaccination

At the discretion of the investigator, any antibiotic, antiviral, anti-pneumosporic, antifungal or other preventive measure

Medicaments for the prophylaxis and treatment of IRR, including antihistamines, antipyretics, corticosteroids, bronchodilators and other medicaments (according to the medical indications)

Agents for preventing and treating TLS, including uric acid lowering agents (e.g., allopurinol, labyrine), diuretics, and phosphate binders and potassium binders. Labyrinthine is contraindicated in participants with baseline G6PD deficiency (for further information on labyrinthine, reference is made to the pharmaceutical instructions or SmPC)

Non-live SARS-CoV-2 vaccine authorized or approved by researchers

Allowing for treatment of active SARS-CoV-2 infection or high risk exposure, including using experimental therapies and taking into account the risk of DDI (e.g., pa Luo Weide), and should be discussed with medical inspectors.

Preventive measures allowing the use of anticoagulant or antiplatelet agents are decided by researchers. Researchers are recommended to carefully assess the potential risk of thromboembolism and hemorrhage for individual participants.

Any supplementary medication, such as vitamins and mineral supplements, is allowed.

Subject to the notes described in section 7.7.1, any concomitant medication or therapy deemed necessary for the care of the participant should be used. Repeated PK evaluations may be performed if changes are made to concomitant medications suspected of affecting drug absorption or metabolism.

7.7.3 Other limitations and precautions. The ability of the study intervention to induce photosensitivity has not been fully evaluated. Participants receiving study intervention must be offered consultation to minimize exposure to ultraviolet light, wear protective clothing and sunglasses when out in the sun, and use broad spectrum Ultraviolet (UV) a/UVB sunscreens with a sun protection factor of at least 25 on their exposed skin until at least 2 days after permanent discontinuation of study intervention.

If the use of an antiarrhythmic agent known to prolong QT interval and/or other agents known to prolong QT interval is necessary, the risk and benefit should be discussed with the sponsor's medical inspector before use with research intervention.

7.7.3.1 Imaging limitations and precautions. The local imaging facility is responsible for determining the appropriate imaging modality and imaging protocol for each imaging study based on participant attributes (e.g., allergy history, diabetes history, renal status). Imaging contraindications and contrast risks should be considered in this evaluation. It will be assessed whether a participant suffering from renal insufficiency should receive contrast media and, if so, which contrast media and dosage are appropriate. Particularly for Magnetic Resonance Imaging (MRI), participants with severe renal insufficiency (i.e., estimated glomerular filtration rate [ eGFR ] < 30 mL/min/1.73 m 2) are at increased risk of nephrogenic systemic fibrosis, and therefore MRI contrast agents are contraindicated. Furthermore, if the participants have tattoos, metal implants, pacemakers, etc., they may be excluded from MRI. These limitations will be outlined in the imaging handbook.

Mild hydration before and after IV contrast agents should follow local standard care. The final decision to perform MRI on individual participants who entered this study depends on the study center radiologist, the researcher, and criteria set by the local ethics committee.

7.8 Study intervention was continued after study termination. At the end of the study, if the study intervention is not available as an approved treatment in the local country, the participant who continues to exhibit clinical benefit will be eligible to receive the sponsor-supplied study intervention for the longest treatment duration as specified in section 7.1. If research interventions are not available as approved and available treatments, they will be provided via extension of the study, rolling studies requiring approval by the institutional health and ethics committee, or by another mechanism decided by the sponsor.

The sponsor retains the right to terminate the study intervention supplied by the sponsor if any, a) the study is terminated due to a safety issue, b) the development of BMS-986458 is terminated for other reasons, including but not limited to lack of efficacy and/or failure to meet the study objective, or c) the participant may obtain a medication from a government sponsor or other health program. In all cases, the sponsor will follow the local regulations.

8. The criteria are aborted. Details of the study at a particular study center or discontinuation of the study as a whole are provided.

8.1 Discontinuation of study intervention. The participants must discontinue IP/IMP (and non-IMP/AxMP as determined by the investigator) for any of the following reasons:

participants were asked to stop study intervention. Participants who require discontinuation of study intervention will remain in the study and should continue to follow-up with respect to the protocol-prescribed follow-up procedure. The only exception to this is when the participant explicitly withdraws consent to any further contact with the person he/she or the participant previously authorized to provide this information.

Researchers believe that continued participation in the study does not meet any clinical AEs, laboratory test result abnormalities, or intermittent conditions of optimal interest to the participants.

The sponsor terminates the study.

The ability to freely provide consent is lost due to being trapped or involuntarily curbed for the treatment of mental or physical disorders (e.g., infectious diseases). ( Note that in certain situations and only in countries where local regulations allow, caged participants may be allowed to continue as participants. Stringent conditions apply and require sponsor approval. )

As described in section 7.4.1, any event that results in a delay in dosing for > 4 weeks.

In rare cases, participants may benefit from recovering BMS-986458 after an interruption longer than 4 weeks. All decisions about whether to resume administration after > 4 weeks of disruption must be discussed with the medical inspector.

Evidence of clinically significant disease progression based on the judgment of the investigator.

Pregnancy (reference section 9.2.5).

Significant non-compliance with the regimen (e.g., procedure, assessment, medication, etc.). Researchers should discuss such problems with medical inspectors.

For data collected at treatment discontinuation and follow-up, and any further evaluation that may be completed, refer to section 2: activity schedule.

All participants who discontinued the study intervention should follow the follow-up procedure as prescribed by the protocol outlined in section 2: activity schedule. The only exception to this requirement is when the participants withdraw consent to all study procedures (including post-treatment study follow-up), or lose the ability to freely consent (e.g., trapped or involuntarily banned for treatment of mental or physical illness).

If study intervention is discontinued before the participants complete the study, the reasons for the discontinuation must be recorded in the participant's medical record and entered on the corresponding eCRF page according to the local regulatory requirements of each region/country.

8.1.1 Study follow-up after study intervention. Participants who discontinued study intervention may continue to be followed. The BMS may require that survival data be collected for all treated participants outside of the protocol-defined window. At this request, each participant will be contacted to determine its state of survival unless the participant has withdrawn consent or no visit for all contacts.

8.2 Discontinuing the study

The participants should inform the investigator of the decision to withdraw consent from future follow-up.

Withdrawal consent should be interpreted in detail by the investigator in the medical record to indicate whether to withdraw from further treatment with study intervention alone or withdraw from study procedure and/or study follow-up after treatment, and entered on the appropriate eCRF page.

If a life state is being measured (whether the participant is alive or dead), the life state should be determined using publicly available information only as directed appropriately according to local law.

If the participant withdraws consent disclosing future information, the sponsor can retain and continue to use any data collected prior to this withdrawal of consent.

8.2.1 Individual suspension criteria

The participant may either withdraw from the study entirely at any time, according to his/her own requirements, or withdraw from the study at any time as determined by the researcher for safety, behavioral, compliance or regulatory reasons. This is not expected to be common. Stopping study intervention is not considered a withdrawal from the study.

When the study is discontinued, an early termination visit should be made, if possible, as shown in section 2: activity schedule. For data collected at study discontinuation and follow-up, and any further evaluation that needs to be completed, see activity schedule.

At that time, the participants will permanently discontinue both study intervention and study.

8.3 Lose access. If the participant fails to return to the clinic for the required study visit, the following must be taken:

All reasonable effort must be made to locate participants to determine and report their persistence status. This includes follow-up of persons authorized by the participant.

A missed visit is defined as an inability to reach the participant after a minimum of 3 recorded telephone, fax or email messages, and a lack of a participant's reply to a 1-note mailing message. All attempts should be recorded in the participant's medical record.

If the researcher uses a third party representative to assist in the follow-up portion of the study that has been included in the participant's informed consent, the researcher may use a sponsor-retained third party representative to assist the research center staff in obtaining participant contact information or other public life status data, such as public health registries and databases, required to complete the follow-up portion of the study.

Research center staff and representatives will consult publicly available sources, such as public health registries and databases, to obtain updated contact information.

If the participant remains out of visit after all attempts, the last known survival date as determined by the researcher should be reported and recorded in the participant's medical record.

If it is determined that the participant has died, the research center will use locally allowed methods to obtain the date and cause of the death.

9. Study evaluation and procedure

Study procedure and time settings are summarized in section 2: activity schedule.

No scheme exemption or exemption is allowed.

It must be immediately discussed with the sponsor when all direct safety issues occur or are recognized to determine if the participant should continue or discontinue treatment.

Adherence to study design requirements, including those specified in the activity schedule, is critical to and essential to study implementation.

All screening evaluations must be completed and reviewed prior to group entry to confirm that the potential participants meet all qualification criteria. The researcher will maintain a screening log to record details of all participants screened and confirm qualifications or record reasons for screening failure (if applicable).

For screening or baseline purposes, procedures performed as part of the participants' routine clinical management (e.g., whole blood count) and procedures obtained prior to signing informed consent can be utilized, provided that the procedures meet the criteria defined by the protocol and have been performed within the timeframe defined in the activity schedule.

The maximum blood volume collected from each participant (including any additional assessment that may be needed) will not exceed 920 mL over the duration of the study.

Repeated or unscheduled samples may be collected for safety reasons or technical problems with the sample. Additional measurements are made, including laboratory tests that are not required for research, according to clinical indications or compliance with local regulations.

Some of the evaluations mentioned in this section may not be collected as data in eCRF. They are intended for use as safety monitors by the treating physician. Additional tests or evaluations may be performed as clinically needed or as required by institutional or local regulations.

9.1 Efficacy assessment

9.1.1 Efficacy evaluation of the study. The evaluation of the antitumor activity of BMS-986458 will be determined by the investigator. Efficacy assessment ("rukino standard") will be performed using malignant lymphoma international working group standard ²⁰ and multi-dimensional standard （Deauville Criteria for fluorodeoxyglucose-positron emission tomography (FDG-PET) scan interpretation）^29,30 explained by fluorodeoxyglucose-positron emission tomography (FDG-PET) scan.

Efficacy assessment included the following:

Clinical findings (e.g., physical examination, systemic symptoms)

Contrast enhanced Computed Tomography (CT) scanning and FDG-PET or FDG-PET-CT where appropriate

Bone marrow examination (bone marrow biopsy [ BMB ] and bone marrow aspiration [ BMA ]) where appropriate

Magnetic Resonance Imaging (MRI) where appropriate

Screening tumor assessments would include the following:

CT scan with contrast agent CT is the recommended imaging modality. Contrast enhanced MRI of the abdomen and pelvis is also permissible. Standard care CT scans or MRI performed up to 56 days prior to cycle 1 day 1 may be acceptable for screening, even before ICF signing.

All participants will be subjected to baseline FDG-PET or FDG-PET/CT scans, except those with known FDG-PET negative tumors.

Only participants with known/suspected bone marrow involvement were subjected to bone marrow aspiration and biopsy.

Brain/spinal scans were performed only on participants with suspected brain involvement (if clinical indications were present) (contrast enhanced MRI is preferred, whereas CT may be performed if MRI is contraindicated).

In-treatment tumor assessment will be performed as follows:

Using the same scanning modality as at screening, radiological assessment will be performed as noted in the activity schedule (section 2) until the researcher assesses progress, withdrawal of consent, death, or longest study duration. If the previous scan is within 28 days, then an EOT scan need not be obtained. If study intervention is discontinued for reasons other than disease progression, imaging should continue every 26 weeks from the last dose for up to 52 weeks or until disease progression, death, start of new anti-cancer therapy, failure to visit, withdrawal of consent from the study, or end of the study, whichever occurs first.

For participants with bone marrow involvement at the time of screening, bone marrow assessment by flow immunophenotyping will be performed to confirm Complete Response (CR). All participants discontinuing treatment for reasons other than disease progression or recurrence, starting new anti-cancer therapy, or withdrawal of consent from the overall study will follow-up every 6 months (+ -14 days) until progression, starting new systemic anti-cancer therapy including SCT, or withdrawal, for up to 1 year after treatment is completed. After progression, CT scanning is no longer required according to the protocol, however, evaluation can continue according to local clinical practice.

Tumor response at the time of evaluation after each screening will be determined by based on the investigator's revised criteria for malignant lymphoma response. Treatment decisions will be based on the assessment of the investigator. ²⁰

Survival will be determined approximately every 12 weeks after disease progression for up to 1 year. Anticancer therapies including systemic treatment, cell therapy, SCT, surgery and radiation will be collected after disease progression.

9.1.2 Imaging evaluation of the study. Images for tumor assessment will be submitted to a central imaging provider for collection in a research central repository and may undergo Blind Independent Central Review (BICR) at any time during and/or after the study. Before scanning the first participant, the study center should be qualified and understand the image acquisition guidelines and submission process as outlined in the imaging manual provided by the center imaging provider. Any additional imaging (including scanning at unscheduled time points and/or at an external institution) that may exhibit tumor response or progression for response assessment by the rupulos standard ²⁰ must be collected and submitted to a central imaging provider. For further details, refer to section 2. Clinical data may be provided to the imaging provider if applicable.

A slice thickness of 5mm or less should be used and the image acquired without an intermediate gap (continuous). For all imaging time points, all efforts should be made to image each participant on the same scanner using the same acquisition protocol. Where possible, tumor measurements should be made by the same researcher or radiologist for each evaluation.

In the presence of contraindications of contrast agents or MRI (e.g. incompatible pacemakers), the following guidelines are strongly recommended:

if the participant has contraindications for CT intravenous contrast agent, contrast agent-free CT of the chest should be obtained as well as contrast agent enhanced MRI of the abdomen, pelvis and other known/suspected disease sites.

If the participant has contraindications of both MRI and CT intravenous contrast agent, contrast agent free CT of the chest should be obtained as well as contrast agent free MRI of the abdomen, pelvis and other known/suspected disease sites.

If the participant has a contraindication for MRI in addition to the contraindication of CT intravenous contrast agent, contrast agent-free CT of chest, abdomen, pelvis and other known/suspected disease sites is acceptable.

The low dose or attenuation-corrected CT portions of combined PET-CT have limited use in efficacy assessment and therefore it is suggested that they should not replace dedicated diagnostic enhancers to enhance CT scans for rugasan measurements. However, if the research center is able to have records that indicate that the CT performed as part of the PET-CT has the same diagnostic quality as the diagnostic CT (with IV and oral contrast), then the CT part of the PET-CT can be used for the rugasan measurement.

9.2 Adverse events. Definition of AE or SAE can be found in appendix 3.

The AE will be reported by the participant (or caretaker or agent, as appropriate).

Researchers and any qualified prescribers are responsible for detecting, recording and reporting events that meet the definition of AE or SAE, and continue to be responsible for follow-up of AEs that are serious, considered relevant to study intervention or study, or that lead to participants discontinuing the study prior to completion of the study. For SAE report, refer to appendix 3.

Safety reporting of all AEs and SAE on eCRF will be performed using CTCAE v5.0 definition and fractionation, except for the oncolytic reaction (NCI CTCAE V3.0.0) and oncolytic syndrome. ²⁶

9.2.1 Time period and frequency of collecting AE and SAE information. All AEs and SAE had to be collected from signing the informed consent form (including those considered relevant to the program prescribed by the protocol) and until 28 days after discontinuation of dosing or last scheduled visit as noted in the activity schedule (section 2).

The researcher must report any SAE that occurs after these periods of time and is considered relevant to the research intervention or protocol-prescribed procedure (e.g., follow-up skin biopsies).

All SAE will be recorded and reported to sponsors or designated personnel in time and not more than 24 hours after the event is known, as indicated in annex 3.

Researchers will submit any updated SAE data to sponsors or designated personnel within 24 hours of the time that updated information is available.

The investigator had no obligation to actively look for AE or SAE of the study participants before. However, if the researcher learns of any SAE (including death) at any time after the participant exits the study and he/she thinks that the event is reasonably relevant to the study intervention or study participation, the researcher must timely inform the sponsor.

Methods of evaluating and assessing the causal relationship of AE and SAE and procedures for completing and reporting/communicating SAE reports are provided in appendix 3.

9.2.2 Methods for detecting AE and SAE. AE may report spontaneously or be initiated during open interrogation, examination or evaluation of participants. Care should not be taken to introduce bias in collecting AE and/or SAE. Where appropriate for a project or regimen, interrogation of a particular AE should be directed by clinical judgment with known AEs.

9.2.3 Follow-up for AE and SAE

All SAE will be followed until resolved, until the condition stabilizes, until the event is otherwise interpreted, or until the participant is out of visit (as defined in section 8.3: out of visit).

Non-severe AEs should be followed to regress or stabilize, or reported as SAE if severe (see appendix 3).

Follow-up (as the case may be) is also required for non-severe AEs that lead to disruption or cessation of study intervention, as well as those that occur at the end of study intervention.

All identified non-severe AEs must be recorded and described on the non-severe AE page (paper or electronic version) of the CRF. Supplementation of CRF may need to be accomplished for AE and/or laboratory test results reported/identified during the study.

Further information about the follow-up procedure is given in appendix 3.

9.2.5 Pregnancy. If a participant is found to be pregnant or likely to have pregnant at the time of study intervention exposure (including at least 28 days after study intervention administration) after study intervention begins, the researcher must immediately notify the sponsor medical inspector/prescriber of the event and complete the pregnancy monitoring form within 24 hours of learning of the event and forward to the sponsor prescriber. According to the reporting procedure described in appendix 3, the pregnancy report must follow the same transmission time settings and procedures reported to the BMS as those used for SAE.

In all cases, study intervention will be discontinued in an appropriate manner (e.g., dose taper if necessary for safety of the participants).

Follow-up information about the pregnancy process, including perinatal and neonatal outcome (as applicable) offspring information, must be reported on the pregnancy monitoring tables. Procedures for study discontinuation and follow-up protocol requirements must be performed.

Any pregnancy occurring in the female partner of a male participant at the time of study intervention exposure (including at least 28 days after study intervention administration) should be reported to the sponsor or to the prescribing personnel. For a sponsor or prescribing person to collect any pregnancy monitoring information from a female partner, the female partner must sign an ICF disclosing this information. Information about this pregnancy will be collected and recorded on a pregnancy monitoring table.

According to the reporting procedure described in appendix 3, any pregnant individual (e.g. caregivers, pharmacists, study coordinator, inspector) exposure to BMS-986458 must follow the same transmission time settings and procedures reported to BMS as those for SAE.

If any sexual activity involving penile intercourse (e.g., vaginal, anal, oral) occurs between a male participant and one or more pregnancy partners without the use of a condom during and at least 28 days after administration of the study intervention, the sponsor or prescribing person should be notified of the information even if the male participant has undergone successful vasectomy.

9.2.6 The laboratory test results are abnormal. The following laboratory test result anomalies should be collected on the adverse event-Non-severe event and severe event eCRF (ADVERSE EVENTS-Non-serious and Serious EVENTS ECRF) page. The paper form is only intended to be a back-up option when the electronic system is not functioning.

Any laboratory test results clinically significant or conforming to the SAE definition

Any laboratory test outcome anomalies requiring participant discontinuation or discontinuation of study intervention

Any laboratory test outcome abnormalities requiring participants to receive specific corrective therapies

It is contemplated that where possible, report researchers will use clinical terms rather than laboratory terms (e.g., anemia rather than low hemoglobin values).

9.2.7 Potential drug-induced liver injury. Specific criteria for identifying potential drug-induced liver injury (DILI) for this protocol have not been identified. Standard medical practice for identifying and monitoring liver problems should be followed.

9.2.8 Other security considerations. Any significant worsening of the condition noted during mid-term or final physical examination, ECG, radiographic imaging, or any other potential safety assessment that is required or not required by the protocol should also be recorded as a non-severe AE or SAE (as applicable), and reported accordingly.

9.3 Overdosing. Overdosing is defined as the accidental or intentional administration of any dose of the product, which is considered both overdosing and medically important. Overdosing, which meets regulatory definition of SAE, will be reported as SAE (see appendix 3).

If overdosing occurs, the researcher/treating physician should:

Immediate contact with the medical inspector.

Closely monitoring participants for AE/SAE and laboratory test result anomalies,

The amount of overdose was recorded in eCRF and the duration of overdose.

9.4 Safety. The planned time points for all security assessments are listed in section 2, the activity schedule. Starting after cycle 6, it was decided by the investigator that clinical visits and evaluations on day 1 could be conducted on a less frequent schedule in the absence of any ≡2 toxicity associated with the study intervention.

9.4.1 Physical examination and ECOG physical performance status. Physical examinations will be performed, including height (screening only and EoT), weight, BMI and BSA (when applicable), and ECOG physical status. If PE screening is performed within 24 hours prior to day 1 dosing, a single examination can be counted as both screening and pre-dosing evaluation. Any findings during the PE will be recorded as a medical history in the participant's medical record and on the appropriate eCRF.

9.4.2 Vital signs. Vital signs include body temperature, blood pressure, heart rate, and pulse oximetry monitoring. Blood pressure and heart rate should be measured after the participants rest quietly for at least 5 minutes. According to the evaluation of the researcher, additional vital signs should be obtained according to the clinical indications. The recorded measurements will be collected in the participant's medical record and on the appropriate eCRF.

9.4.3 An electrocardiogram. ECG collection time points were repeated in triplicate for part a and part B, respectively. After the participant has been in the supine or semi-prone position for at least 10 minutes, a 12-lead ECG (12 leads at 25 mm/sec, reported heart rate, heart rate/ventricular rate, PR interval, QRS complex, QT interval, and QT interval corrected using the Fridericia formula [ QTcF ]). Triplicate 12-lead ECGs should be collected at each time point using sponsor-supplied machines (3 recordings at 2±1 minute intervals) and sent to a central ECG laboratory for third party evaluation. At the end of each visit, triplicate ECG outputs are uploaded to a central ECG laboratory for deterministic analysis and interpretation. The researcher will immediately make a clinical decision based on its interpretation of the safety ECG results.

During the treatment period, specific triplicate ECGs will be collected within about 15 minutes prior to the time-matched PK sample.

Thereafter, if there is a clinical indication after cycle 4, a single ECG will be performed. If after cycle 4, then only one ECG is needed for the EOT visit. The ECG starting at cycle 5 will be a single ECG and will not be sent to the central ECG laboratory. Clinically significant changes to these ECGs will be recorded in the AE section of eCRF.

9.4.4 ECHO or MUGA scans. LVEF will be determined by multi-gated acquisition scans or echocardiography at screening and EOT. Follow-up assessment should be performed based on clinical indications. The follow-up assessment should use the same procedure as used in the screening assessment. Clinically significant decreases are defined as > 20% absolute decreases or decreases to below 45% relative to baseline LVEF.

9.4.5 Clinical safety laboratory assessment. Researchers must record their reviews of each laboratory security report. Clinical safety laboratory tests will be performed in a local laboratory that will provide a reference range for these tests. The results of safety clinical laboratory tests performed during screening and at dosing visits must be available and reviewed prior to dosing.

Table B4-9.4.5-1 clinical laboratory evaluation

Abbreviations ALT (SGPT) alanine aminotransferase (serum glutamate pyruvate aminotransferase), aPTT, activated partial thromboplastin time, AST (SGOT), aspartate aminotransferase (serum glutamate oxaloacetate aminotransferase), BUN, blood urea nitrogen, CBC, whole blood count, HIV, human immunodeficiency virus, ig, immunoglobulin, INR, international normalized ratio, LDH, lactate dehydrogenase, PT, prothrombin time, WOCBP, females with fertility potential.

9.4.6 Other security assessments. Any occasional findings of potential clinical relevance not directly related to the protocol goals should be evaluated and processed by the investigator of the study according to standard medical/clinical judgment.

9.4.7 Pregnancy test. Women with fertility potential (WOCBP) are defined as sexually mature women, having:

No hysterectomy or double sided ovariectomy or double sided tubal resection was performed.

After not reaching natural menopause (amenorrhea after anticancer therapy does not exclude fertility potential) for at least 12 consecutive months (e.g. having menopause at any time in the preceding 12 consecutive months).

The investigator would classify female participants as WOCBP according to this definition. WNOCBP participants do not need pregnancy tests, but must record the reason in the eCRF and source files. For WOCBP, a medical supervised pregnancy test with a minimum sensitivity of 25 mIU/mL has to be performed. Pregnancy tests will be performed by a local laboratory.

For WOCBP, pregnancy tests will be performed at the visits listed below and are detailed in PPP. Serum pregnancy tests were obtained at screening (10 to 14 days before 1 st day of cycle 1) and another pregnancy test (serum or urine) had to be performed 24 hours before 1 st day of study treatment. Participants had to receive BMS-986458 until the investigator verified that 2 screening pregnancy tests were negative.

Serum or urine pregnancy tests should be performed within 24 hours prior to day 1 of each subsequent cycle and at the end of treatment (EOT) visit. Participants had to receive BMS-986458 until the investigator verified that the pregnancy test was negative.

WOCBP with regular or no menstrual cycle must agree to conduct pregnancy tests weekly during the first 28 days of study participation and then every 28 days when BMS-986458 is taken, at the end of the study, and monthly for at least 28 days after the last dose of BMS-986458.

WOCBP with irregular menstrual cycles must agree to conduct pregnancy tests weekly for the first 28 days of study participation and then every 14 days when BMS-986458 is taken, at the end of the study, every 14 days at least 28 days after the last dose of BMS-986458.

Male participants WOCBP or their companion WOCBP must avoid activity that could lead to pregnancy for at least 28 days after the last dose of BMS-986458.

Pregnancy test results will be recorded in the source file and eCRF.

9.5 Pharmacokinetics. Where the data allows, the pharmacokinetics of BMS-986458 and its diastereomers will be derived from plasma concentrations and time data during monotherapy and combination, as shown below.

If the data allows, the PK parameters to be evaluated (as applicable) include, but are not limited to, the following:

Individual participant PK parameter values will be derived by a validated PK analysis procedure by a non-atrioventricular method. The actual time will be used for analysis.

PK data from this study can be combined with PK data from other studies to characterize PK using population PK analysis. The results of this analysis will be reported separately (see section 10.4: statistical analysis for additional details).

9.5.1 Pharmacokinetic sampling instructions and collection windows. Blood samples will be collected from all participants at the selected visit.

It is expected that all efforts will be made to collect PK samples at the times indicated in the PK sampling schedule. The exact sampling date and time should be recorded accurately at all times. An illustration of PK sample collection is provided below:

Samples prior to dosing will be taken within minutes prior to oral dosing. All mid-treatment time points are intended to be consistent with the date of administration of study intervention. If the dose is known to be delayed, a pre-dose sample should be collected just prior to the delayed dose. However, if a pre-dose sample is collected but the dose is delayed, no additional pre-dose sample should be collected.

9.5.2 Pharmacokinetic sample analysis. Plasma will be used to determine the concentration of BMS-986458 diastereomers (BMT-648592 and BMT-648599) using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Plasma BMS-986458 concentrations will be derived from BMT-648592 and BMT-648599 concentrations. Exploratory LC-MS/MS methods (as applicable) can be used to conduct exploratory analysis of BMS-986458 metabolites in plasma using blood samples collected for PK evaluation.

Treatment distribution will be distributed to bioanalytical laboratories to minimize unnecessary analysis and/or re-analysis of PK study samples.

Bioanalytical samples designated for evaluation (e.g., PK, biomarker) from the same collection time point can be used interchangeably for analysis, if desired.

In addition, the residual bioassay samples will be archived for up to 20 years after the end of the study or for the longest period of time allowed by applicable law, and may be used for potential exploratory bioassays (including but not limited to metabolite assays, etc.) and/or for additional method purposes (including but not limited to cross-validation, PK selectivity, critical points, etc.).

9.7 Biomarkers

9.7.1 Germ line genetics. Blood samples will be collected from all participants for DNA isolation unless local laws/regulations prohibit or if not approved by IRB/IEC. In these cases, the participants may be instructed about DNA collection as optional collection. For biomarker sampling schedules, refer to tables 2-5 in section 2, activity schedules.

The collection and transport information for these samples is in the laboratory/procedure manual. The sponsor stores the DNA sample in a secure storage space with sufficient measures to protect confidentiality. Unless banned by local laws/regulations or IRB/IEC, samples may remain beyond the end of the study for up to 20 years to understand the genetic mechanism of the response and/or the disease/disorder and related conditions under study.

9.7.3 Tumor biopsies. Tumor biopsies are mandatory for all participants in part a and part B, except when the tumor is not reachable (or in a location where the participants may be at risk), the researcher will discuss with the medical inspector the possibility of omitting the in-treatment biopsy.

Biopsies were collected by tumor resection or core needle (4 punctures recommended) at the following time points:

Screening biopsies

O is only done after qualification has been determined by all other criteria:

resecting or core needle fresh biopsies from one or more sites no earlier than 2 weeks after discontinuing the previous anti-cancer therapy.

O only after discussion with sponsor medical inspectors, the investigator may still be charged to the group of participants who consider no disease sites that can be safely biopsied percutaneously or by other minimally invasive techniques:

Instead of screening for fresh biopsies, diagnostic archive FFPEs (tumor mass or slice/slab specimens) can be submitted if the following criteria are met:

if it was obtained no more than 1 year before the group was entered, collected after the last intervention and only after a discussion with sponsor medical inspectors.

If there are sufficient materials available for pharmacodynamic evaluation. For detailed information, refer to the laboratory manual.

Biopsy during treatment

O for participants in part B only, tumor biopsies were taken in one forced treatment at cycle 1 on days 13, 14 or 15.

O in treatment (optional). If an in-treatment biopsy is collected during clinical care in either part a or part B, an optional sample may be collected from the in-treatment biopsy with a paired blood sample for research purposes.

End of treatment biopsy (optional)

Optimally, tumor tissue samples will be obtained from the same tumor site.

Biopsies should be treated as detailed in the laboratory manual. Fine needle aspirate is not sufficient as a source of tumor biopsy material. The anatomic pathology report should be sent with the biopsy material.

9.7.4 Fine needle aspirate (part a only). All participants in part a will need to collect FNA samples for pharmacodynamic biomarker studies. This sample will be collected at the time of screening (only if an archive sample is submitted instead of screening for tumor biopsies) and at cycle 1, day 13, 14 or 15. If fresh tumor biopsies are collected at the time of screening, the submitted portion of the sample is used for pharmacodynamic assessment by flow cytometry instead of FNA.

If the tumor is not accessible (or at a location that would put the participant at risk), the researcher will discuss with the medical inspector the possibility of omitting the FNA or replacing the FNA in the treatment with a core needle biopsy (core biopsy).

9.8 Optionally future studies. This protocol will include residual sample storage for optional future study. Samples reserved for future studies will be stored in sponsor-designated storage facilities. The manager of these samples will ensure that they are used correctly throughout their lifetime and will destroy the samples at the end of the scheduled storage period (not more than 20 years after the end of the study or the longest period allowed by applicable law). The sample will be stored in coded form and no key will be available to the researcher. The key will be held securely by the researchers at the clinical research center and therefore the researchers have no direct ability to associate the sample with a particular individual.

10. Statistical considerations

10.1 Statistical assumptions. BMS-986458 administered as a single agent or in combination with an anti-lymphoma agent would be expected to exhibit sufficient safety and tolerability, as well as beneficial benefit/risk profiles to support further clinical testing. No formal evaluation of the prospective hypothesis was made.

10.2 And determining the sample size. The sample size is considered as follows:

For monotherapy dose escalation in part A1, no formal statistical efficacy calculations to determine sample size will be performed. The actual number depends on the number of DLTs observed, the number of effective responses, and the number of dosage levels required to determine one or more OBDs (optimal biological dosages) to select RP2D based on the phase-2 phase I design (phase 1: BOIN, phase 2: TITE-BOIN). We used this 2-phase design to adequately evaluate toxicity, also to evaluate early efficacy, and did not delay participant entry into the group and treatment during our dose escalation/decrementing period. If we encounter unexpectedly high toxicity at the first dose level we will decrement to dose level-1, otherwise dose escalation of dose levels 2 to 5 will be considered. The expected number of participants in phase 1 and phase 2 that could evaluate toxicity and primary efficacy was between 3 and 16 per dose level, about 41 participants from both phase 1 and phase 2 of part A1 (appendix 8 and appendix 9; fig. 13A and 13B). BOIN designs merely direct the phase 1 increment/decrement decision based on toxicity. If BOIN directed dose escalation i) reaches and confirms the highest dose level within acceptable toxicity or ii) finds an MTD based on a pre-specified range of toxicity, phase 1 (BOIN) will enter phase 2 (TITE-BOIN). Additional participants of phase 2 will then be assigned in the allowed doses defined in section 10.4.2.1 based on the TITE-BOIN design. Up to 16 participants (together with participants from stage 1) will be treated at each dose level to select OBD in stage 2.

For the combination with rituximab in part A2, the starting dose of BMS-986458 will be determined by SRC based on the emerging safety, PK and pharmacodynamic data from part A1 and part B1 (as defined in section 5.1.2.1). Up to 16 participants will be treated per dose level, with up to 32 total participants rated for response. Dose escalation from lower combined dose to higher combined dose was performed based on the same BOIN criteria as phase 1 in part A1.

In parts B1 and B2, 2 OBDs will be studied per indication to further confirm tolerability and early efficacy of selected OBDs of single agent (part A1) and combination (part A2). For monotherapy and combination, each indication and dose level will be grouped with a sample size of approximately 20 participants that can evaluate the response to further evaluate safety and assess primary efficacy in parts B1 and B2. In DLBCL, 55% of the observed CRR or 11 of 20 participants will have a bilateral 95% Crepper-Pearson confidence interval (CI; 31.5%, 76.9%), the lower limit of which will be higher than the CRR 28% ³¹ as reported in the RE-MIND2 study. In FL, 17 of the 85% observed CRR or 20 participants had CR that would have a bilateral 95% Crepper-Pearson confidence interval (CI; 62.1%, 96.8%), the lower limit of which would be higher than CRR 60% ³² as reported in Bartlett et al (2022). These expansion queues will also allow further observation of the preliminary efficacy signal under OBD, which will guide the selection of RP 2D.

For these expansion cohorts and participants who were treated at the same dose level in dose escalation, we will use bayesian continuous monitoring to determine early cessation of rules-treatment at potentially toxic dose levels that may not be fully observed was paused or suspended during dose escalation studies in parts A1 and A2. Safety information from all subjects treated at these dose levels (including both up-dosing and up-dosing cohorts) collected until the end of cycle 4 will be included in the continuous toxicity monitoring. For participants who will meet DLT criteria (except for dose reduction due to AEs that do not meet DLT criteria), the excess toxicity rate (based on DLT definition) will be monitored. We will use as an early cessation rule the statistical criteria that the upper limit probability of toxicity is exceeded based on a posterior probability of less than 60% (excess toxicity based on DLT > 0.3|data, a, b) > = 0.6, where a and b are parameters of the a priori distribution of β (a, b) for the DLT rate hypothesis: 0.30 for the target toxicity rate, a=0.3, b=0.7 (see appendix 10 for its operating characteristics and continuous toxicity monitoring boundaries), and will update the a priori to derive the posterior (e.g. if 0 DLT is observed in 6 participants at a certain dose level, the updated posterior will be β [0.3, 6.7 ]) taking into account the number of DLTs observed at the selected dose level.

10.3 The collection is analyzed. For analytical purposes, the following populations were defined:

10.4 Statistical analysis

10.4.1 General considerations apply. Statistical Analysis Plans (SAPs) will be formulated and finalized before database locking and will describe the choice of participants to be included in the analysis and the procedure taking into account missing, unused and spurious data. The following is a summary of the planned statistical analysis of the primary endpoint and the secondary endpoint.

10.4.2 One or more primary endpoints

TABLE B4-10.4.2-1 primary endpoint

Abbreviations AE, adverse events, CTCs, general term criteria, DLT, dose limiting toxicity, PT, preferred terms, SAE, serious adverse events.

Table B4-10.4.2.-2 summary of the main measured attributes of the main endpoint

Abbreviations AE, adverse events, DLT, dose limiting toxicity, NHL, non-Hodgkin's lymphoma, R/R, recurrent/refractory, SAE, severe adverse events

10.4.2.1 Dose up/down rule. A 2-stage dose escalation design will be employed in part A1. The first stage designs ^33,34 to find the MTD based on Bayesian Optimal Intervals (BOIN). In stage 1, BOIN designs are implemented (see appendix 8). Target toxicity rate was q > =0.30. Participants will be grouped into a size 3 queue and treated therein. DLTs are defined in section 7.4.4 and only those DLTs occurring during the first period will be used for dose discovery.

If BOIN directed dose escalation i) reaches and confirms the highest dose level within acceptable toxicity or ii) finds an MTD based on a pre-specified range of toxicity, phase 1 (BOIN) will enter phase 2 (TITE-BOIN). At stage 2, the dose escalation/decrementing will follow TITE-BOIN12 trial design ^35•36, where the target tolerability toxicity (DLT rate) is 30%, the lowest baseline acceptable work efficiency (ORR) of DLBCL and FL is 30%, and the efficacy score reflects the clinical desirability of toxicity and efficacy outcomes (i.e., risk-benefit tradeoff). TITE-BOIN are extensions of BOIN12 to mitigate delayed toxicity and efficacy data. Only TITE features were used to interpolate less than 50% of efficacy data based on efficacy posterior distribution, while toxicity data availability was required to be complete (100%), as was BOIN for toxicity assessment. When the study goes from stage 1 (BOIN) to stage 2 (TITE-BOIN), it is expected that at least 50% or more of the participant efficacy data will likely be available, so the study can proceed without too much delay. However, if at least 50% of efficacy data for a certain dose level has not been collected at the end of phase 1, a moderate suspension study is expected to define the next dose level for OBD discovery under the TITE-BOIN design. Table 10.4.2.1-1 shows the utility attributed to each possible efficacy toxicity outcome. To protect the participants from toxic and/or ineffective doses, two dose acceptability criteria were used. A dose is considered to be adoptable and eligible for treatment of a participant if it meets the following safety and efficacy criteria based on the TITE-BOIN guidelines.

(Safety) Pr (DLT Rate > 0.3I data) < 0.8, (efficacy) Pr (ORR < 0.30I data) < 0.75

The unadopted dose will be eliminated from the trial. If a dose is eliminated due to toxicity, all doses above that dose will also be eliminated.

TABLE B4-10.4.2.1-1 Utility Table for Utility score for TITE-BOIN12 design

10.4.3 One or more secondary endpoints. Statistical analysis of the secondary endpoint schedule is summarized in table 10.4.3-1.

TABLE B4-10.4.3-1 secondary endpoint

Abbreviations AUC, area under concentration-time curve, cmax, maximum concentration, DOR, duration of reaction, KM, kaplan-Meyer, ORR, total reaction rate, OS, total lifetime, PFS, progression free lifetime, PK, pharmacokinetics, RR, reaction rate, tmax, time to maximum observed concentration, TTR, time to reaction.

TABLE 10.4.3-2 summary of the major measured attributes of key secondary objectives

Abbreviations AE, adverse events, BICR, blind independent central examination, BOR, optimal overall response, CI, confidence interval, CR, complete response, DLT, dose limiting toxicity, ORR, total response rate, PR, partial response, RECIST, response evaluation criteria for solid tumors, SAE, severe adverse events.

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³⁵ Lin R, Zhou Y, Yan F, et al. BOIN12: Bayesian Optimal Interval Phase I/II Trial Design for Utility-Based Dose Finding in Immunotherapy and Targeted Therapies. JCO Precision Oncology 2020; 4; 1393-1402.

³⁶ Zhou Y, Lin R, Lee J, et al. TITE-BOIN12: A Bayesian Phase I/II Trial Design to Find the Optimal Biological Dose with Late-onset Toxicity and Efficacy. Stat. Med. 2022; 41; 1918- 1931.

Appendix B4-1 abbreviation

Appendix B4-3 definition and procedure of adverse events and serious adverse events: recording, evaluation, follow-up and reporting

Adverse events

SAE definition. If the event is not an AE according to the definition above, it cannot be an SAE even if severe conditions are met.

Serious adverse events

Pregnancy and DILI must follow the same delivery time settings and procedures to sponsors as those used for SAE. (for reporting pregnancy, see section 9.2.5 in the protocol: pregnancy.)

Evaluation of AE and SAE

Reporting SAE to sponsors or designated personnel

SAE (whether or not related to study intervention) and pregnancy must be reported to sponsors (or designated personnel) in time and not more than 24 hours after the informed event.

SAE must be recorded on SAE report form.

The claimed method for SAE data reporting is performed by means of an electronic case report form (eCRF).

The o-paper SAE report form is intended only as a back-up option when the electronic data collection system is not available/able to transmit eCRF to the sponsor (or designated person).

In this case, the paper form is transmitted via an email or an acknowledged facsimile transmission.

When paper forms are used, the original paper forms should remain in place.

Pregnancy must be recorded on a paper pregnancy monitoring form and transmitted via email or confirmed fax transmission.

Appendix B4-8 flow and operational characteristics of BOIN design in part A1 and part A2, part 1

For the purpose of overdose control, if Pr (pj > 0.30|data) > 0.95 and at least 3 evaluable participants received treatment at dose level j, where pj is the true DLT rate at dose level j (j=1,..5), dose j and higher will be eliminated from further examination. The posterior probability is evaluated based on a beta-binomial model of yj I pj-binomial distribution (pj), where pj-uniform distribution (0, 1), where yj is the number of participants experiencing DLT at dose level j. When the lowest dose was eliminated, the trial was stopped for safety. The probability threshold of 0.95 is chosen to be consistent with common practice, i.e., to eliminate the dose of 2/3 of the participants experiencing DLT when the target DLT rate is less than or equal to 1/3. A simulation study was performed with 5000 simulation trials, with a queue size of 3 and a maximum of 10 queues. The target toxicity was set to 0.3 and the trial was stopped if the number of patients allocated to each dose reached 9. The operating characteristics of this dose up/down rule are presented in the table above.

Appendix B4-9 flow and operational characteristics of the TITE-BOIN design in section A1, stage 2

As with the phase 2 design in part A1, this design will begin with the most dose selected in phase 1.

The proposed TITE-BOIN design will use bayesian interpolation to supplement missing or unobserved missing efficacy data (if the missing is less than 50%). Bayesian interpolation is one of the most stringent statistical methods for processing delayed efficacy data. Therefore, the missing efficacy data will be bayesian data supplemented based on the posterior distribution as follows. Assuming that the joint outcome Y follows the Dirichlet-polynomial model (Dirichlet-multinomial model), when YT and YE deletions are observed, we extract the missing values of YE from their conditional posterior. A pre-specified a priori setting (a01=4/8, a00=1/8, a11=2/8, a10=1/8) will be used for (YT, YE) = (01, 00, 11, 10), which a priori setting will prefer higher doses (similar to the traditional RP2D discovery based on MTD), but once efficacy data of > 50% (n > =2) is available, its effect will diminish rapidly. Following interpolation, BOIN algorithm can be applied directly to guide dose conversion and identification of OBD. A simulation study was performed with 5000 simulation trials, with a queue size of 3 and a maximum of 10 queues. The target toxicity was set to 0.3 and the trial was stopped if the number of patients allocated to each dose reached 10. The DLT evaluation window was 1 month, the efficacy evaluation window was 2 months, the accumulation rate was 3 patients/month, if 50% or more of the efficacy data were pending, we would halt accumulation, and the safety and efficacy criteria and utility table were set as described in section 10.4.2.1. The starting dose in phase 2 in the simulation is set to the dose selected the most in phase 1, in particular doses 2,3, 5, 1 of scenes 1-5. The operating characteristics of the TITE-BOIN design are provided in the table above.

Appendix B4-10 continuous toxicity monitoring plans in extended queues

Continuous toxicity monitoring will be performed on subjects treated with the dose selected for the dose expansion cohort during the dose expansion and dose escalation phases. During dose extension (part B), non-binding bayesian safety monitoring methods will be used to continuously monitor for excessive toxicity, defined as the occurrence of AEs meeting the DLT criteria defined in dose escalation. Weak information priors, e.g. beta (0.3, 0.7), will be constructed for estimating the stopping boundary. After a minimum of 6 subjects were enrolled and at least one cycle of study treatment was completed, the following unconstrained bayesian stop criteria were calculated to halt the trial based on the posterior distribution of excessive toxicity.

Probability (excessive toxicity > 0.3|data, a=0.3, b=0.7) > =0.60,

Where a=0.3 and b=0.7 is a parameter of the β prior distribution.

This criterion means that when the probability of toxicity exceeding 0.3 is greater than 0.60, the study can be paused. The excess toxicity rate of 0.3 was determined to be consistent with the target toxicity rate specified in the dose escalation. The operational characteristics of developing a bayesian safety monitoring plan (part B) at a specified period are detailed in the table below based on bayesian toxicity monitoring analysis tools at triadesign.

Although the invention has been described in detail by way of illustration and example for the purpose of clarity of understanding, the description and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific documents cited herein are expressly incorporated by reference in their entirety.

Claims

1. A method of treating cancer or an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of an anti-CD 20 antibody and a compound of formula (IA):

(IA)

or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

2. A method of treating cancer or an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (IA):

(IA)

or a pharmaceutically acceptable salt thereof, wherein:

R ^1a and R ^1b are each independently H, halo or C ₁-C₆ alkyl;

Or R ^2a and R ^2b together form oxo;

Or R ^1a and R ^2a together form a bridging C ₂-C₃ alkylene;

R ^3a and R ^3b are each independently H, halo or C ₁-C₆ alkyl;

R ^4a and R ^4b are each independently H, halo or C ₁-C₆ alkyl,

R ^5a and R ^5b are each independently H, halo or C ₁-C₆ alkyl;

w is 0 or 1;

R ⁶ is H or C ₁-C₆ alkyl;

R ⁷ is H or C ₁-C₆ alkyl;

x and y are each independently 0 or 1, provided that x and y are not both 1;

r ⁸ is Cl or-CN;

r ⁹ is F;

X ³ is N or CH;

z is 0 or 1;

r ^10a and R ^10b are each independently H or halo;

Wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O and S;

r ¹² is H, halo or C ₁-C₆ alkyl;

r ¹³ is H or halo;

R ¹⁴ is H or C ₁-C₆ alkyl;

x ⁴ is N or CR ¹⁵;

R ¹⁵ is H or C ₁-C₆ alkyl;

X ⁵ and X ⁶ are each independently N or CH, and

Is a single bond or a double bond;

wherein one or more hydrogen atoms in the compound are optionally replaced with deuterium,

Wherein the compound of formula (IA) or a pharmaceutically acceptable salt thereof is administered (i) daily on each day of a 28-day cycle, or (ii) on days 1-14 of a 28-day cycle and not on days 15-28 of the 28-day cycle, or (iii) on days 1-21 of a 28-day cycle and not on days 22-28 of the 28-day cycle, or

Wherein the compound of formula (IA) or a pharmaceutically acceptable salt thereof is administered to the subject once daily or twice daily at a dose of about 20 mg/day, about 40 mg/day, about 80 mg/day, about 160 mg/day, about 260 mg/day, about 400 mg/day, or about 640 mg/day.

3. The method of claim 1 or 2, wherein the compound of formula (IA) or a pharmaceutically acceptable salt thereof is administered to the subject once daily or twice daily at a dose of about 20 mg/day, about 40 mg/day, about 80 mg/day, about 160 mg/day, about 260 mg/day, about 400 mg/day, or about 640 mg/day.

4. The method of any one of claims 1-3, wherein the compound of formula (IA), or a pharmaceutically acceptable salt thereof, is administered (i) daily on each day of a 28-day cycle, or (ii) on days 1-14 of a 28-day cycle and not on days 15-28 of the 28-day cycle, or (iii) on days 1-21 of a 28-day cycle and not on days 22-28 of a 28-day cycle.

5. The method of any one of claims 2-4, further comprising administering an anti-CD 20 antibody to the subject.

6. The method of claim 1 or 5, wherein the anti-CD 20 antibody is rituximab.

7. The method of claim 6, wherein rituximab is administered intravenously.

8. The method of claim 6 or 7, wherein about 375 mg/m ² rituximab is administered once every 28 days.

9. The method of any one of claims 6-8, wherein rituximab is administered on day 1 of a 28-day treatment cycle and the compound of formula (IA) or a pharmaceutically acceptable salt thereof is administered (i) daily on each day of the 28-day treatment cycle, or (ii) on days 1-14 of the 28-day treatment cycle and not on days 15-28 of the 28-day treatment cycle, or (iii) on days 1-21 of the 28-day treatment cycle and not on days 22-28 of the 28-day treatment cycle.

10. The method of any one of claims 1-9, wherein the method is a method of treating cancer.

11. The method of claim 10, wherein the cancer is lymphoma.

12. The method of claim 11, wherein the lymphoma is B-cell lymphoma.

13. The method of claim 11 or 12, wherein the lymphoma is non-hodgkin's lymphoma.

14. The method of any one of claims 11-13, wherein the lymphoma is diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, marginal zone lymphoma, burkitt's lymphoma, or lymphoplasmacytic lymphoma.

15. The method of claim 14, wherein the lymphoma is diffuse large B-cell lymphoma or follicular lymphoma.

16. The method of any one of claims 10-15, wherein the cancer is recurrent or refractory.

17. The method of any one of claims 1-16, wherein the compound has the formula:

(70),

or an isomer thereof, or a pharmaceutically acceptable salt of the foregoing.

18. The method of claim 17, wherein the compound has the formula:

(70),

or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition comprising a compound of the formula:

(70),

20. A unit dose formulation comprising the pharmaceutical composition of claim 19, wherein the compound (70) or an isomer thereof or a pharmaceutically acceptable salt of the foregoing is present in the unit dosage form in an amount of from about 10 mg to about 100 mg.